214 results on '"Gye Won Han"'
Search Results
2. Structural insights on ligand recognition at the human leukotriene B4 receptor 1
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Nairie Michaelian, Anastasiia Sadybekov, Élie Besserer-Offroy, Gye Won Han, Harini Krishnamurthy, Beata A. Zamlynny, Xavier Fradera, Phieng Siliphaivanh, Jeremy Presland, Kerrie B. Spencer, Stephen M. Soisson, Petr Popov, Philippe Sarret, Vsevolod Katritch, and Vadim Cherezov
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Science - Abstract
Human leukotriene B4 receptors (BLT1 and BLT2) are members of the GPCR superfamily that respond to a potent pro-inflammatory lipid and chemoattractant LTB4. Here authors determined a crystal structure of the human BLT1 in complex with a selective antagonist MK-D-046 and provide insights into hBLT1 ligand recognition and its mechanism of action.
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- 2021
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- View/download PDF
3. Harnessing the power of an X-ray laser for serial crystallography of membrane proteins crystallized in lipidic cubic phase
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Ming-Yue Lee, James Geiger, Andrii Ishchenko, Gye Won Han, Anton Barty, Thomas A. White, Cornelius Gati, Alexander Batyuk, Mark S. Hunter, Andrew Aquila, Sébastien Boutet, Uwe Weierstall, Vadim Cherezov, and Wei Liu
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g-protein-coupled receptors ,membrane proteins ,xfels ,serial femtosecond crystallography ,adenosine a2a receptors ,lipidic cubic phases ,high dynamic range detectors ,Crystallography ,QD901-999 - Abstract
Serial femtosecond crystallography (SFX) with X-ray free-electron lasers (XFELs) has proven highly successful for structure determination of challenging membrane proteins crystallized in lipidic cubic phase; however, like most techniques, it has limitations. Here we attempt to address some of these limitations related to the use of a vacuum chamber and the need for attenuation of the XFEL beam, in order to further improve the efficiency of this method. Using an optimized SFX experimental setup in a helium atmosphere, the room-temperature structure of the adenosine A2A receptor (A2AAR) at 2.0 Å resolution is determined and compared with previous A2AAR structures determined in vacuum and/or at cryogenic temperatures. Specifically, the capability of utilizing high XFEL beam transmissions is demonstrated, in conjunction with a high dynamic range detector, to collect high-resolution SFX data while reducing crystalline material consumption and shortening the collection time required for a complete dataset. The experimental setup presented herein can be applied to future SFX applications for protein nanocrystal samples to aid in structure-based discovery efforts of therapeutic targets that are difficult to crystallize.
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- 2020
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- View/download PDF
4. Full-length human GLP-1 receptor structure without orthosteric ligands
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Fan Wu, Linlin Yang, Kaini Hang, Mette Laursen, Lijie Wu, Gye Won Han, Qiansheng Ren, Nikolaj Kulahin Roed, Guangyao Lin, Michael A. Hanson, Hualiang Jiang, Ming-Wei Wang, Steffen Reedtz-Runge, Gaojie Song, and Raymond C. Stevens
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Science - Abstract
Glucagon-like peptide-1 receptor (GLP-1R) plays an important role in glucose homeostasis and treatment of type 2 diabetes. Here authors report the peptide-free crystal structure of human GLP-1R in an inactive state which reveals a unique closed conformation of the extracellular domain.
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- 2020
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5. Structural basis of ligand binding modes at the human formyl peptide receptor 2
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Tong Chen, Muya Xiong, Xin Zong, Yunjun Ge, Hui Zhang, Mu Wang, Gye Won Han, Cuiying Yi, Limin Ma, Richard D. Ye, Yechun Xu, Qiang Zhao, and Beili Wu
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Science - Abstract
Formyl peptide receptors (FPRs) are GPCRs that play important roles in transducing chemotactic signals in phagocytes and mediating host-defense and inflammatory responses. Here the authors present the 2.8 Å crystal structure of human FPR2 in complex with the peptide agonist WKYMVm and in combination with molecular docking, ligand-binding and signalling assays provide further insights into the binding modes of FPR2 to both non-formyl and formyl peptides.
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- 2020
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- View/download PDF
6. Structural basis of ligand selectivity and disease mutations in cysteinyl leukotriene receptors
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Anastasiia Gusach, Aleksandra Luginina, Egor Marin, Rebecca L. Brouillette, Élie Besserer-Offroy, Jean-Michel Longpré, Andrii Ishchenko, Petr Popov, Nilkanth Patel, Taku Fujimoto, Toru Maruyama, Benjamin Stauch, Margarita Ergasheva, Daria Romanovskaia, Anastasiia Stepko, Kirill Kovalev, Mikhail Shevtsov, Valentin Gordeliy, Gye Won Han, Vsevolod Katritch, Valentin Borshchevskiy, Philippe Sarret, Alexey Mishin, and Vadim Cherezov
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Science - Abstract
Cysteinyl leukotriene G protein-coupled receptors CysLT1 and CysLT2 regulate pro-inflammatory responses associated with allergic disorders. Here, authors describe four crystal structures of CysLT2R in complex with three dual CysLT1R/CysLT2R antagonists, which shed light on CysLTR ligand selectivity.
- Published
- 2019
- Full Text
- View/download PDF
7. Toward G protein-coupled receptor structure-based drug design using X-ray lasers
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Andrii Ishchenko, Benjamin Stauch, Gye Won Han, Alexander Batyuk, Anna Shiriaeva, Chufeng Li, Nadia Zatsepin, Uwe Weierstall, Wei Liu, Eriko Nango, Takanori Nakane, Rie Tanaka, Kensuke Tono, Yasumasa Joti, So Iwata, Isabel Moraes, Cornelius Gati, and Vadim Cherezov
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drug discovery ,g protein-coupled receptors ,serial femtosecond crystallography ,x-ray free-electron lasers ,structure determination ,membrane proteins ,protein structure ,molecular recognition ,Crystallography ,QD901-999 - Abstract
Rational structure-based drug design (SBDD) relies on the availability of a large number of co-crystal structures to map the ligand-binding pocket of the target protein and use this information for lead-compound optimization via an iterative process. While SBDD has proven successful for many drug-discovery projects, its application to G protein-coupled receptors (GPCRs) has been limited owing to extreme difficulties with their crystallization. Here, a method is presented for the rapid determination of multiple co-crystal structures for a target GPCR in complex with various ligands, taking advantage of the serial femtosecond crystallography approach, which obviates the need for large crystals and requires only submilligram quantities of purified protein. The method was applied to the human β2-adrenergic receptor, resulting in eight room-temperature co-crystal structures with six different ligands, including previously unreported structures with carvedilol and propranolol. The generality of the proposed method was tested with three other receptors. This approach has the potential to enable SBDD for GPCRs and other difficult-to-crystallize membrane proteins.
- Published
- 2019
- Full Text
- View/download PDF
8. Molecular Mechanism for Ligand Recognition and Subtype Selectivity of α2C Adrenergic Receptor
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Xiaoyu Chen, Yueming Xu, Lu Qu, Lijie Wu, Gye Won Han, Yu Guo, Yiran Wu, Qingtong Zhou, Qianqian Sun, Cenfeng Chu, Jie Yang, Liu Yang, Quan Wang, Shuguang Yuan, Ling Wang, Tao Hu, Houchao Tao, Yaping Sun, Yunpeng Song, Liaoyuan Hu, Zhi-Jie Liu, Raymond C. Stevens, Suwen Zhao, Dong Wu, and Guisheng Zhong
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Biology (General) ,QH301-705.5 - Abstract
Summary: Adrenergic G-protein-coupled receptors (GPCRs) mediate different cellular signaling pathways in the presence of endogenous catecholamines and play important roles in both physiological and pathological conditions. Extensive studies have been carried out to investigate the structure and function of β adrenergic receptors (βARs). However, the structure of α adrenergic receptors (αARs) remains to be determined. Here, we report the structure of the human α2C adrenergic receptor (α2CAR) with the non-selective antagonist, RS79948, at 2.8 Å. Our structure, mutations, modeling, and functional experiments indicate that a α2CAR-specific D206ECL2-R409ECL3-Y4056.58 network plays a role in determining α2 adrenergic subtype selectivity. Furthermore, our results show that a specific loosened helix at the top of TM4 in α2CAR is involved in receptor activation. Together, our structure of human α2CAR-RS79948 provides key insight into the mechanism underlying the α2 adrenergic receptor activation and subtype selectivity. : Chen et al. report the crystal structure of human α2CAR and the functional experimental results, which indicate that extracellular regions determine α2 adrenergic subtype selectivity. The structural and functional results provide the molecular explanation for α2CAR selective ligands and insights to understand GPCR subtype selectivity. Keywords: α2C adrenergic receptor, subtype selectivity, crystal structure, GPCRs, Raynaud's syndrome, JP1302
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- 2019
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9. Structural Basis of the Diversity of Adrenergic Receptors
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Lu Qu, Qingtong Zhou, Yueming Xu, Yu Guo, Xiaoyu Chen, Deqiang Yao, Gye Won Han, Zhi-Jie Liu, Raymond C. Stevens, Guisheng Zhong, Dong Wu, and Suwen Zhao
- Subjects
Biology (General) ,QH301-705.5 - Abstract
Summary: Adrenergic receptors are highly homologous while at the same time display a wide diversity of ligand and G-protein binding, and understanding this diversity is key for designing selective or biased drugs for them. Here, we determine two crystal structures of the α2A adrenergic receptor (α2AAR) in complex with a partial agonist and an antagonist. Key non-conserved residues from the ligand-binding pocket (Phe7.39 and Tyr6.55) to G-protein coupling region (Ile34.51 and Lys34.56) are discovered to play a key role in the interplay between partial agonism and biased signaling of α2AAR, which provides insights into the diversity of ligand binding and G-protein coupling preference of adrenergic receptors and lays the foundation for the discovery of next-generation drugs targeting these receptors. : Crystal structures of α2A adrenergic receptor (α2AAR) reveal the molecular basis for the diversity in adrenergic receptors. Qu et al. define compelling roles for key amino acids in ligand binding, partial agonism, and biased signaling of α2AAR. Keywords: GPCR, α2A adrenergic receptor, partial agonism, biased signaling
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- 2019
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10. Crystal structure of a multi-domain human smoothened receptor in complex with a super stabilizing ligand
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Xianjun Zhang, Fei Zhao, Yiran Wu, Jun Yang, Gye Won Han, Suwen Zhao, Andrii Ishchenko, Lintao Ye, Xi Lin, Kang Ding, Venkatasubramanian Dharmarajan, Patrick R. Griffin, Cornelius Gati, Garrett Nelson, Mark S. Hunter, Michael A. Hanson, Vadim Cherezov, Raymond C. Stevens, Wenfu Tan, Houchao Tao, and Fei Xu
- Subjects
Science - Abstract
Smoothened receptors (SMO) play a key role in the Hedgehog signalling pathway. Here the authors present the structure of a multi-domain human SMO with a rationally designed stabilizing ligand bound in the transmembrane domain of the receptor, and propose a model for SMO activation.
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- 2017
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11. Structural insights into the molecular mechanisms of myasthenia gravis and their therapeutic implications
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Kaori Noridomi, Go Watanabe, Melissa N Hansen, Gye Won Han, and Lin Chen
- Subjects
myasthenia gravis ,nicotinic acetylcholine receptors ,crystallography ,autoantibodies ,Pichia pastoris ,Medicine ,Science ,Biology (General) ,QH301-705.5 - Abstract
The nicotinic acetylcholine receptor (nAChR) is a major target of autoantibodies in myasthenia gravis (MG), an autoimmune disease that causes neuromuscular transmission dysfunction. Despite decades of research, the molecular mechanisms underlying MG have not been fully elucidated. Here, we present the crystal structure of the nAChR α1 subunit bound by the Fab fragment of mAb35, a reference monoclonal antibody that causes experimental MG and competes with ~65% of antibodies from MG patients. Our structures reveal for the first time the detailed molecular interactions between MG antibodies and a core region on nAChR α1. These structures suggest a major nAChR-binding mechanism shared by a large number of MG antibodies and the possibility to treat MG by blocking this binding mechanism. Structure-based modeling also provides insights into antibody-mediated nAChR cross-linking known to cause receptor degradation. Our studies establish a structural basis for further mechanistic studies and therapeutic development of MG.
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- 2017
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12. GPCR Agonist-to-Antagonist Conversion: Enabling the Design of Nucleoside Functional Switches for the A2A Adenosine Receptor
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Anna Shiriaeva, Daejin Park, Gyudong Kim, Yoonji Lee, Xiyan Hou, Dnyandev B. Jarhad, Gibae Kim, Jinha Yu, Young Eum Hyun, Woomi Kim, Zhan-Guo Gao, Kenneth A. Jacobson, Gye Won Han, Raymond C. Stevens, Lak Shin Jeong, Sun Choi, and Vadim Cherezov
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Drug Discovery ,Molecular Medicine - Published
- 2022
13. Structural insight into apelin receptor-G protein stoichiometry
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Yang Yue, Lier Liu, Li-Jie Wu, Yiran Wu, Ling Wang, Fei Li, Junlin Liu, Gye-Won Han, Bo Chen, Xi Lin, Rebecca L. Brouillette, Émile Breault, Jean-Michel Longpré, Songting Shi, Hui Lei, Philippe Sarret, Raymond C. Stevens, Michael A. Hanson, and Fei Xu
- Subjects
Apelin Receptors ,GTP-Binding Proteins ,Structural Biology ,Humans ,Carrier Proteins ,Molecular Biology ,Receptors, G-Protein-Coupled ,Signal Transduction - Abstract
The technique of cryogenic-electron microscopy (cryo-EM) has revolutionized the field of membrane protein structure and function with a focus on the dominantly observed molecular species. This report describes the structural characterization of a fully active human apelin receptor (APJR) complexed with heterotrimeric G protein observed in both 2:1 and 1:1 stoichiometric ratios. We use cryo-EM single-particle analysis to determine the structural details of both species from the same sample preparation. Protein preparations, in the presence of the endogenous peptide ligand ELA or a synthetic small molecule, both demonstrate these mixed stoichiometric states. Structural differences in G protein engagement between dimeric and monomeric APJR suggest a role for the stoichiometry of G protein-coupled receptor- (GPCR-)G protein coupling on downstream signaling and receptor pharmacology. Furthermore, a small, hydrophobic dimer interface provides a starting framework for additional class A GPCR dimerization studies. Together, these findings uncover a mechanism of versatile regulation through oligomerization by which GPCRs can modulate their signaling.
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- 2022
14. Structural basis of GABA reuptake inhibition
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Zenia Motiwala, Nanda Gowtham Aduri, Hamidreza Shaye, Gye Won Han, Jordy Homing Lam, Vsevolod Katritch, Vadim Cherezov, and Cornelius Gati
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GABA Plasma Membrane Transport Proteins ,Neurotransmitter Agents ,Multidisciplinary ,Protein Conformation ,Cryoelectron Microscopy ,Humans ,Anticonvulsants ,GABA Uptake Inhibitors ,Tiagabine ,gamma-Aminobutyric Acid ,Article - Abstract
γ-Aminobutyric acid (GABA) transporter 1 (GAT1)(1) regulates neuronal excitation of the central nervous system by clearing the synaptic cleft of the inhibitory neurotransmitter GABA upon its release from synaptic vesicles. Elevating the levels of GABA in the synaptic cleft, by inhibiting GABA reuptake transporters, is an established strategy to treat neurological disorders, such as epilepsy(2). Here we determined the cryo-electron microscopy structure of full-length, wild-type human GAT1 in complex with its clinically used inhibitor tiagabine(3), with an ordered part of only 60 kDa. Our structure reveals that tiagabine locks GAT1 in the inward-open conformation, by blocking the intracellular gate of the GABA release pathway, and thus suppresses neurotransmitter uptake. Our results provide insights into the mixed-type inhibition of GAT1 by tiagabine, which is an important anticonvulsant medication. Its pharmacodynamic profile, confirmed by our experimental data, suggests initial binding of tiagabine to the substrate-binding site in the outward-open conformation, whereas our structure presents the drug stalling the transporter in the inward-open conformation, consistent with a two-step mechanism of inhibition(4). The presented structure of GAT1 gives crucial insights into the biology and pharmacology of this important neurotransmitter transporter and provides blueprints for the rational design of neuromodulators, as well as moving the boundaries of what is considered possible in single-particle cryo-electron microscopy of challenging membrane proteins.
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- 2022
15. The activation mechanism and antibody binding mode for orphan GPR20
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Xi Lin, Shan Jiang, Yiran Wu, Xiaohu Wei, Gye-Won Han, Lijie Wu, Junlin Liu, Bo Chen, Zhibin Zhang, Suwen Zhao, Vadim Cherezov, and Fei Xu
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Genetics ,Cell Biology ,Molecular Biology ,Biochemistry - Abstract
GPR20 is a class-A orphan G protein-coupled receptor (GPCR) and a potential therapeutic target for gastrointestinal stromal tumors (GIST) owing to its differentially high expression. An antibody-drug conjugate (ADC) containing a GPR20-binding antibody (Ab046) was recently developed in clinical trials for GIST treatment. GPR20 constitutively activates Gi proteins in the absence of any known ligand, but it remains obscure how this high basal activity is achieved. Here we report three cryo-EM structures of human GPR20 complexes including Gi-coupled GPR20 in the absence or presence of the Fab fragment of Ab046 and Gi-free GPR20. Remarkably, the structures demonstrate a uniquely folded N-terminal helix capping onto the transmembrane domain and our mutagenesis study suggests a key role of this cap region in stimulating the basal activity of GPR20. We also uncover the molecular interactions between GPR20 and Ab046, which may enable the design of tool antibodies with enhanced affinity or new functionality for GPR20. Furthermore, we report the orthosteric pocket occupied by an unassigned density which might be essential for exploring opportunities for deorphanization.
- Published
- 2023
16. Inhibition of the angiotensin II type 2 receptor AT
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Richard, Perryman, Alexander, Renziehausen, Hamidreza, Shaye, Androniki D, Kostagianni, Antonis D, Tsiailanis, Thomas, Thorne, Maria V, Chatziathanasiadou, Gregory B, Sivolapenko, Mohamed Ahmed, El Mubarak, Gye Won, Han, Barbara, Zarzycka, Vsevolod, Katritch, Guillaume, Lebon, Cristiana, Lo Nigro, Laura, Lattanzio, Sophie V, Morse, James J, Choi, Kevin, O'Neill, Zoi, Kanaki, Apostolos, Klinakis, Tim, Crook, Vadim, Cherezov, Andreas G, Tzakos, and Nelofer, Syed
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Protein Conformation, alpha-Helical ,Analgesics ,Brain Neoplasms ,Angiotensin II ,Drug Repositioning ,Humans ,Apoptosis ,Angiotensin II Type 2 Receptor Blockers ,Benzhydryl Compounds ,Glioblastoma ,Isoquinolines ,Receptor, Angiotensin, Type 2 ,Tumor Burden - Abstract
Glioblastoma (GBM) is an aggressive malignant primary brain tumor with limited therapeutic options. We show that the angiotensin II (AngII) type 2 receptor (AT
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- 2023
17. Inhibition of the angiotensin II type 2 receptor AT 2 R is a novel therapeutic strategy for glioblastoma
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Richard Perryman, Alexander Renziehausen, Hamidreza Shaye, Androniki D. Kostagianni, Antonis D. Tsiailanis, Thomas Thorne, Maria V. Chatziathanasiadou, Gregory B. Sivolapenko, Mohamed Ahmed El Mubarak, Gye Won Han, Barbara Zarzycka, Vsevolod Katritch, Guillaume Lebon, Cristiana Lo Nigro, Laura Lattanzio, Sophie V. Morse, James J. Choi, Kevin O’Neill, Zoi Kanaki, Apostolos Klinakis, Tim Crook, Vadim Cherezov, Andreas G. Tzakos, Nelofer Syed, Brain Tumour Research Campaign, Medicinal chemistry, and AIMMS
- Subjects
Analgesics ,renin angiotensin system ,Multidisciplinary ,SDG 3 - Good Health and Well-being ,glioblastoma ,Humans ,Apoptosis ,angiotensin II ,Receptor, Angiotensin, Type 2 ,Receptor, Angiotensin, Type 1 - Abstract
Glioblastoma (GBM) is an aggressive malignant primary brain tumor with limited therapeutic options. We show that the angiotensin II (AngII) type 2 receptor (AT 2 R) is a therapeutic target for GBM and that AngII, endogenously produced in GBM cells, promotes proliferation through AT 2 R. We repurposed EMA401, an AT 2 R antagonist originally developed as a peripherally restricted analgesic, for GBM and showed that it inhibits the proliferation of AT 2 R-expressing GBM spheroids and blocks their invasiveness and angiogenic capacity. The crystal structure of AT 2 R bound to EMA401 was determined and revealed the receptor to be in an active-like conformation with helix-VIII blocking G-protein or β-arrestin recruitment. The architecture and interactions of EMA401 in AT 2 R differ drastically from complexes of AT 2 R with other relevant compounds. To enhance central nervous system (CNS) penetration of EMA401, we exploited the crystal structure to design an angiopep-2–tethered EMA401 derivative, A3E. A3E exhibited enhanced CNS penetration, leading to reduced tumor volume, inhibition of proliferation, and increased levels of apoptosis in an orthotopic xenograft model of GBM.
- Published
- 2022
18. Cryo-EM structure of constitutively active human Frizzled 7 in complex with heterotrimeric Gs
- Author
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Bo Chen, Fei Xu, Michel Bouvier, Shane C. Wright, Ainoleena Turku, Chao Li, Lu Xu, Maria Kowalski-Jahn, Paweł Kozielewicz, Yiran Wu, Gye Won Han, Carl-Fredrik Bowin, Hannes Schihada, Gunnar Schulte, and Xianjun Zhang
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Models, Molecular ,Frizzled ,Protein Conformation ,Cryo-electron microscopy ,Constitutively active ,Oncogenes ,Cell Biology ,Biology ,Cell biology ,Cryoelectron microscopy ,Heterotrimeric G protein ,Humans ,Letter to the Editor ,Molecular Biology - Published
- 2021
19. Harnessing the power of an X-ray laser for serial crystallography of membrane proteins crystallized in lipidic cubic phase
- Author
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Alexander Batyuk, Gye Won Han, Andrii Ishchenko, Mark S. Hunter, Sébastien Boutet, Vadim Cherezov, Andrew Aquila, Uwe Weierstall, Cornelius Gati, Wei Liu, Anton Barty, Thomas A. White, James Geiger, and Ming-Yue Lee
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0301 basic medicine ,serial femtosecond crystallography ,Materials science ,g-protein-coupled receptors ,membrane proteins ,02 engineering and technology ,Biochemistry ,law.invention ,X-ray laser ,03 medical and health sciences ,adenosine a2a receptors ,law ,General Materials Science ,ddc:530 ,lcsh:Science ,Dynamic range ,lipidic cubic phases ,Resolution (electron density) ,Detector ,General Chemistry ,high dynamic range detectors ,021001 nanoscience & nanotechnology ,Condensed Matter Physics ,Laser ,xfels ,Research Letters ,Crystallography ,030104 developmental biology ,Nanocrystal ,Femtosecond ,Vacuum chamber ,lcsh:Q ,0210 nano-technology - Abstract
IUCrJ 7(6), 976 - 984 (2020). doi:10.1107/S2052252520012701, Serial femtosecond crystallography (SFX) with X-ray free-electron lasers (XFELs) has proven highly successful for structure determination of challenging membrane proteins crystallized in lipidic cubic phase; however, like most techniques, it has limitations. Here we attempt to address some of these limitations related to the use of a vacuum chamber and the need for attenuation of the XFEL beam, in order to further improve the efficiency of this method. Using an optimized SFX experimental setup in a helium atmosphere, the room-temperature structure of the adenosine A2A receptor (A2AAR) at 2.0 Å resolution is determined and compared with previous A2AAR structures determined in vacuum and/or at cryogenic temperatures. Specifically, the capability of utilizing high XFEL beam transmissions is demonstrated, in conjunction with a high dynamic range detector, to collect high-resolution SFX data while reducing crystalline material consumption and shortening the collection time required for a complete dataset. The experimental setup presented herein can be applied to future SFX applications for protein nanocrystal samples to aid in structure-based discovery efforts of therapeutic targets that are difficult to crystallize., Published by Chester
- Published
- 2020
20. Structural basis of ligand binding modes at the human formyl peptide receptor 2
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Xin Zong, Cuiying Yi, Hui Zhang, Yechun Xu, Qiang Zhao, Limin Ma, Yunjun Ge, Tong Chen, Beili Wu, Gye Won Han, Muya Xiong, Mu Wang, and Richard D. Ye
- Subjects
0301 basic medicine ,Agonist ,Protein Conformation ,medicine.drug_class ,Science ,General Physics and Astronomy ,Peptide ,Ligands ,Article ,General Biochemistry, Genetics and Molecular Biology ,Formyl peptide receptor 2 ,Structure-Activity Relationship ,03 medical and health sciences ,0302 clinical medicine ,G protein-coupled receptors ,medicine ,Humans ,Amino Acid Sequence ,Receptors, Lipoxin ,lcsh:Science ,Peptide sequence ,X-ray crystallography ,G protein-coupled receptor ,chemistry.chemical_classification ,Binding Sites ,Multidisciplinary ,Formyl peptide receptor ,Chemistry ,Chemotaxis ,General Chemistry ,Ligand (biochemistry) ,Receptors, Formyl Peptide ,Molecular Docking Simulation ,N-Formylmethionine Leucyl-Phenylalanine ,030104 developmental biology ,030220 oncology & carcinogenesis ,Mutation ,Biophysics ,lcsh:Q ,Signal Transduction - Abstract
The human formyl peptide receptor 2 (FPR2) plays a crucial role in host defense and inflammation, and has been considered as a drug target for chronic inflammatory diseases. A variety of peptides with different structures and origins have been characterized as FPR2 ligands. However, the ligand-binding modes of FPR2 remain elusive, thereby limiting the development of potential drugs. Here we report the crystal structure of FPR2 bound to the potent peptide agonist WKYMVm at 2.8 Å resolution. The structure adopts an active conformation and exhibits a deep ligand-binding pocket. Combined with mutagenesis, ligand binding and signaling studies, key interactions between the agonist and FPR2 that govern ligand recognition and receptor activation are identified. Furthermore, molecular docking and functional assays reveal key factors that may define binding affinity and agonist potency of formyl peptides. These findings deepen our understanding about ligand recognition and selectivity mechanisms of the formyl peptide receptor family., Formyl peptide receptors (FPRs) are GPCRs that play important roles in transducing chemotactic signals in phagocytes and mediating host-defense and inflammatory responses. Here the authors present the 2.8 Å crystal structure of human FPR2 in complex with the peptide agonist WKYMVm and in combination with molecular docking, ligand-binding and signalling assays provide further insights into the binding modes of FPR2 to both non-formyl and formyl peptides.
- Published
- 2020
21. High ligand efficiency quinazoline compounds as novel A
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Raphaël, Bolteau, Romain, Duroux, Amélie, Laversin, Brandon, Vreulz, Anna, Shiriaeva, Benjamin, Stauch, Gye Won, Han, Vadim, Cherezov, Nicolas, Renault, Amélie, Barczyk, Séverine, Ravez, Mathilde, Coevoet, Patricia, Melnyk, Maxime, Liberelle, and Saïd, Yous
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Molecular Docking Simulation ,Structure-Activity Relationship ,Purinergic P1 Receptor Antagonists ,Receptor, Adenosine A2A ,Quinazolines ,Ligands ,Adenosine A2 Receptor Antagonists - Abstract
The past fifty years have been marked by the surge of neurodegenerative diseases. Unfortunately, current treatments are only symptomatic. Hence, the search for new and innovative therapeutic targets for curative treatments becomes a major challenge. Among these targets, the adenosine A
- Published
- 2022
22. Toward G protein-coupled receptor structure-based drug design using X-ray lasers
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Uwe Weierstall, Wei Liu, Rie Tanaka, Anna Shiriaeva, Yasumasa Joti, Gye Won Han, Isabel Moraes, Alexander Batyuk, Eriko Nango, Nadia A. Zatsepin, Chufeng Li, So Iwata, Kensuke Tono, Vadim Cherezov, Benjamin Stauch, Takanori Nakane, Cornelius Gati, and Andrii Ishchenko
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serial femtosecond crystallography ,genetic structures ,membrane proteins ,Computational biology ,Biochemistry ,drug discovery ,03 medical and health sciences ,g protein-coupled receptors ,0302 clinical medicine ,Molecular recognition ,Protein structure ,General Materials Science ,protein structure ,lcsh:Science ,030304 developmental biology ,G protein-coupled receptor ,0303 health sciences ,Chemistry ,Drug discovery ,General Chemistry ,equipment and supplies ,Condensed Matter Physics ,Research Papers ,eye diseases ,structure determination ,3. Good health ,x-ray free-electron lasers ,Structure based ,lcsh:Q ,sense organs ,Target protein ,molecular recognition ,030217 neurology & neurosurgery - Abstract
A method is presented for efficient co-crystal structure determination for G protein-coupled receptors taking advantage of serial femtosecond crystallography., Rational structure-based drug design (SBDD) relies on the availability of a large number of co-crystal structures to map the ligand-binding pocket of the target protein and use this information for lead-compound optimization via an iterative process. While SBDD has proven successful for many drug-discovery projects, its application to G protein-coupled receptors (GPCRs) has been limited owing to extreme difficulties with their crystallization. Here, a method is presented for the rapid determination of multiple co-crystal structures for a target GPCR in complex with various ligands, taking advantage of the serial femtosecond crystallography approach, which obviates the need for large crystals and requires only submilligram quantities of purified protein. The method was applied to the human β2-adrenergic receptor, resulting in eight room-temperature co-crystal structures with six different ligands, including previously unreported structures with carvedilol and propranolol. The generality of the proposed method was tested with three other receptors. This approach has the potential to enable SBDD for GPCRs and other difficult-to-crystallize membrane proteins.
- Published
- 2019
23. EXTH-14. INHIBITION OF THE ANGIOTENSIN II TYPE 2 RECEPTOR AT2R IS A NOVEL THERAPEUTIC STRATEGY FOR GLIOBLASTOMA
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Richard Perryman, Alexander Renziehausen, Hamidreza Shaye, Androniki Kostagianni, Antonis Tsiailanis, Tom Thorne, Maria Chatziathanasiadou, Gregory Sivolapenko, Mohamed Ahmed El Mubarak, Gye Won Han, Barbara Zarzycka, Vsevolod Katritch, Guillaume Lebon, Cristiana Lo Nigro, Laura Lattanzio, Sophie Morse, James Choi, Kevin O’Neill, Zoe Kanaki, Apostolos Klinakis, Tim Crook, Vadim Cherezov, Andreas Tzakos, and Nelofer Syed
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Cancer Research ,Oncology ,Neurology (clinical) - Abstract
Glioblastoma (GBM) is a primary malignant brain tumor with poor clinical outcomes. Standard of care consists of surgical debulking followed by radiation and temozolomide, but the tumor invariably recurs, and median survival is only ~18 months. Repurposing drugs used for the treatment of other diseases is a promising avenue to identify novel treatments for this highly aggressive form of cancer. One such class of compounds is angiotensin II (AngII) receptor blockers, commonly used to control blood pressure. We show that ~80% of primary human GBM express the angiotensin II type 2 receptor (AT2R). In the presence of AngII, inhibition of AT2R using either PD123319 or EMA401 significantly inhibits GBM proliferation. This effect was lost in GBM cells with CRISPR/Cas9 mediated knockdown of AT2R. EMA401 inhibited invasion, angiogenesis, reduced GBM spheroid growth and induced apoptosis through caspase 3/7 activation. Furthermore, EMA401 induced changes in a number of growth regulatory pathways including apoptosis, DNA replication and focal adhesion. The crystal structure of AT2R bound to EMA401 revealed the receptor to be in an active-like conformation with helix-VIII blocking G protein or β-arrestin recruitment. We demonstrate that the architecture and interaction of EMA401 with AT2R differs drastically from complexes of AT2R with other compounds. Conjugation of EMA401 to angiopep-2 enhanced its blood brain barrier passage and reduced tumor volume in an orthotopic xenograft model of GBM. Targeting AT2R is a novel therapeutic strategy to treat GBM that should be explored in patients.
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- 2022
24. Allosteric Coupling of Drug Binding and Intracellular Signaling in the A2A Adenosine Receptor
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Matthew T. Eddy, Ming-Yue Lee, Zhan-Guo Gao, Kate L. White, Tatiana Didenko, Reto Horst, Martin Audet, Pawel Stanczak, Kyle M. McClary, Gye Won Han, Kenneth A. Jacobson, Raymond C. Stevens, and Kurt Wüthrich
- Published
- 2021
25. Structural insights on ligand recognition at the human leukotriene B4 receptor 1
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Vadim Cherezov, Gye Won Han, Anastasiia Sadybekov, Nairie Michaelian, Stephen M. Soisson, Jeremy Presland, Élie Besserer-Offroy, Xavier Fradera, Philippe Sarret, Vsevolod Katritch, Kerrie Spencer, Phieng Siliphaivanh, Beata Zamlynny, Petr Popov, and Harini Krishnamurthy
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0301 basic medicine ,Cell type ,Science ,Receptors, Leukotriene B4 ,General Physics and Astronomy ,BLT receptor ,Endogeny ,Computational biology ,Spodoptera ,Crystallography, X-Ray ,Ligands ,General Biochemistry, Genetics and Molecular Biology ,Article ,03 medical and health sciences ,Structure-Activity Relationship ,0302 clinical medicine ,G protein-coupled receptors ,Sf9 Cells ,Animals ,Humans ,Hypoglycemic Agents ,X-ray crystallography ,Multidisciplinary ,Binding Sites ,Chemistry ,Mutagenesis ,Chemotaxis ,Leukotriene B4 Receptor 1 ,General Chemistry ,Ligand (biochemistry) ,Recombinant Proteins ,3. Good health ,Molecular Docking Simulation ,030104 developmental biology ,HEK293 Cells ,Diabetes Mellitus, Type 2 ,Docking (molecular) ,Mutagenesis, Site-Directed ,030217 neurology & neurosurgery - Abstract
The leukotriene B4 receptor 1 (BLT1) regulates the recruitment and chemotaxis of different cell types and plays a role in the pathophysiology of infectious, allergic, metabolic, and tumorigenic human diseases. Here we present a crystal structure of human BLT1 (hBLT1) in complex with a selective antagonist MK-D-046, developed for the treatment of type 2 diabetes and other inflammatory conditions. Comprehensive analysis of the structure and structure-activity relationship data, reinforced by site-directed mutagenesis and docking studies, reveals molecular determinants of ligand binding and selectivity toward different BLT receptor subtypes and across species. The structure helps to identify a putative membrane-buried ligand access channel as well as potential receptor binding modes of endogenous agonists. These structural insights of hBLT1 enrich our understanding of its ligand recognition and open up future avenues in structure-based drug design., Human leukotriene B4 receptors (BLT1 and BLT2) are members of the GPCR superfamily that respond to a potent pro-inflammatory lipid and chemoattractant LTB4. Here authors determined a crystal structure of the human BLT1 in complex with a selective antagonist MK-D-046 and provide insights into hBLT1 ligand recognition and its mechanism of action.
- Published
- 2021
26. A Single Reactive Noncanonical Amino Acid Is Able to Dramatically Stabilize Protein Structure
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Jack C. Li, Fariborz Nasertorabi, Weimin Xuan, Gye Won Han, Raymond C. Stevens, and Peter G. Schultz
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Protein Conformation ,Protein Stability ,Escherichia coli Proteins ,Phenylalanine ,Mutation ,Escherichia coli ,Temperature ,Molecular Medicine ,Homoserine O-Succinyltransferase ,General Medicine ,Crystallography, X-Ray ,Biochemistry ,Article - Abstract
A p-isothiocyanate phenylalanine mutant of the homodimeric protein homoserine o-succinyltransferase (MetA) was isolated in a temperature dependent selection from a library of metA mutants containing noncanonical amino acids. This mutant protein has a dramatic increase of 24 °C in thermal stability compared to the wild type protein. Peptide mapping experiments revealed that the isothiocyanate group forms a thiourea cross-link to the N terminal proline of the other monomer, despite the two positions being30 Å apart in the X-ray crystal structure of the wild type protein. These results show that an expanded set of building blocks beyond the canonical 20 amino acids can lead to significant changes in the properties of proteins.
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- 2019
27. XFEL structures of the human MT2 melatonin receptor reveal the basis of subtype selectivity
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Uwe Weierstall, Reid H.J. Olsen, Raymond C. Stevens, Samuel T. Slocum, Vadim Cherezov, Sahba Zaare, Cornelius Gati, Alexandra R. Tribo, Benjamin Stauch, Chufeng Li, Lan Zhu, Xi Ping Huang, Linda C. Johansson, Gye Won Han, Nadia A. Zatsepin, Saïd Yous, Shuming Hao, Wei Liu, Alexander Batyuk, Bryan L. Roth, John D. McCorvy, Vsevolod Katritch, Jessica M. Grandner, and Nilkanth Patel
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Models, Molecular ,0301 basic medicine ,Mutant ,Electrons ,Molecular Dynamics Simulation ,Ligands ,Melatonin receptor ,Article ,Substrate Specificity ,Melatonin ,Structure-Activity Relationship ,03 medical and health sciences ,0302 clinical medicine ,Extracellular ,medicine ,Humans ,Receptor ,G protein-coupled receptor ,Multidisciplinary ,Receptor, Melatonin, MT2 ,Chemistry ,Lasers ,Receptor, Melatonin, MT1 ,Ligand (biochemistry) ,3. Good health ,Cell biology ,Molecular Docking Simulation ,030104 developmental biology ,Diabetes Mellitus, Type 2 ,Indenes ,Membrane protein ,Mutation ,Crystallization ,030217 neurology & neurosurgery ,medicine.drug - Abstract
The human MT1 and MT2 melatonin receptors1,2 are G-protein-coupled receptors (GPCRs) that help to regulate circadian rhythm and sleep patterns3. Drug development efforts have targeted both receptors for the treatment of insomnia, circadian rhythm and mood disorders, and cancer3, and MT2 has also been implicated in type 2 diabetes4,5. Here we report X-ray free electron laser (XFEL) structures of the human MT2 receptor in complex with the agonists 2-phenylmelatonin (2-PMT) and ramelteon6 at resolutions of 2.8 A and 3.3 A, respectively, along with two structures of function-related mutants: H2085.46A (superscripts represent the Ballesteros–Weinstein residue numbering nomenclature7) and N862.50D, obtained in complex with 2-PMT. Comparison of the structures of MT2 with a published structure8 of MT1 reveals that, despite conservation of the orthosteric ligand-binding site residues, there are notable conformational variations as well as differences in [3H]melatonin dissociation kinetics that provide insights into the selectivity between melatonin receptor subtypes. A membrane-buried lateral ligand entry channel is observed in both MT1 and MT2, but in addition the MT2 structures reveal a narrow opening towards the solvent in the extracellular part of the receptor. We provide functional and kinetic data that support a prominent role for intramembrane ligand entry in both receptors, and suggest that there might also be an extracellular entry path in MT2. Our findings contribute to a molecular understanding of melatonin receptor subtype selectivity and ligand access modes, which are essential for the design of highly selective melatonin tool compounds and therapeutic agents. Structural and functional studies show that the MT2 melatonin receptor, unlike the MT1 receptor, contains an extracellular opening for ligand entry, shedding light on receptor subtype specificity.
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- 2019
28. Structural basis for ligand recognition at the human MT1 melatonin receptor
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Saïd Yous, Wei Liu, Uwe Weierstall, Raymond C. Stevens, Reid H.J. Olsen, Nairie Michaelian, Alexandra R. Tribo, Anna Shiriaeva, Nilkanth Patel, Vadim Cherezov, Xi Ping Huang, Benjamin Stauch, Gye Won Han, Linda C. Johansson, Bryan L. Roth, Cornelius Gati, Vsevolod Katritch, Alexander Batyuk, Caleb Madsen, Samuel T. Slocum, Thomas D. Grant, Jessica M. Grandner, Andrii Ishchenko, W. Brehm, Lan Zhu, John D. McCorvy, and Thomas A. White
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0301 basic medicine ,education.field_of_study ,Multidisciplinary ,Chemistry ,Ligand ,Population ,Article ,3. Good health ,Melatonin ,03 medical and health sciences ,Transmembrane domain ,Pineal gland ,030104 developmental biology ,0302 clinical medicine ,medicine.anatomical_structure ,medicine ,Biophysics ,Serotonin ,Binding site ,education ,Receptor ,030217 neurology & neurosurgery ,medicine.drug - Abstract
Melatonin (N-acetyl-5-methoxytryptamine) is a neurohormone that maintains circadian rhythms1 by synchronization to environmental cues and is involved in diverse physiological processes2 such as the regulation of blood pressure and core body temperature, oncogenesis, and immune function3. Melatonin is formed in the pineal gland in a light-regulated manner4 by enzymatic conversion from 5-hydroxytryptamine (5-HT or serotonin), and modulates sleep and wakefulness5 by activating two high-affinity G-protein-coupled receptors, type 1A (MT1) and type 1B (MT2)3,6. Shift work, travel, and ubiquitous artificial lighting can disrupt natural circadian rhythms; as a result, sleep disorders affect a substantial population in modern society and pose a considerable economic burden7. Over-the-counter melatonin is widely used to alleviate jet lag and as a safer alternative to benzodiazepines and other sleeping aids8,9, and is one of the most popular supplements in the United States10. Here, we present high-resolution room-temperature X-ray free electron laser (XFEL) structures of MT1 in complex with four agonists: the insomnia drug ramelteon11, two melatonin analogues, and the mixed melatonin-serotonin antidepressant agomelatine12,13. The structure of MT2 is described in an accompanying paper14. Although the MT1 and 5-HT receptors have similar endogenous ligands, and agomelatine acts on both receptors, the receptors differ markedly in the structure and composition of their ligand pockets; in MT1, access to the ligand pocket is tightly sealed from solvent by extracellular loop 2, leaving only a narrow channel between transmembrane helices IV and V that connects it to the lipid bilayer. The binding site is extremely compact, and ligands interact with MT1 mainly by strong aromatic stacking with Phe179 and auxiliary hydrogen bonds with Asn162 and Gln181. Our structures provide an unexpected example of atypical ligand entry for a non-lipid receptor, lay the molecular foundation of ligand recognition by melatonin receptors, and will facilitate the design of future tool compounds and therapeutic agents, while their comparison to 5-HT receptors yields insights into the evolution and polypharmacology of G-protein-coupled receptors.
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- 2019
29. Publisher Correction: Structural basis of GABA reuptake inhibition
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Zenia Motiwala, Nanda Gowtham Aduri, Hamidreza Shaye, Gye Won Han, Jordy Homing Lam, Vsevolod Katritch, Vadim Cherezov, and Cornelius Gati
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Multidisciplinary - Published
- 2022
30. Inhibition of the angiotensin II type 2 receptor AT2R is a novel therapeutic strategy for glioblastoma.
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Perryman, Richard, Renziehausen, Alexander, Shaye, Hamidreza, Kostagianni, Androniki D., Tsiailanis, Antonis D., Thorne, Thomas, Chatziathanasiadou, Maria V., Sivolapenko, Gregory B., El Mubarak, Mohamed Ahmed, Gye Won Han, Zarzycka, Barbara, Katritch, Vsevolod, Lebon, Guillaume, Nigro, Cristiana Lo, Lattanzio, Laura, Morse, Sophie V., Choi, James J., O'Neill, Kevin, Zoi Kanaki, and Klinakis, Apostolos
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ANGIOTENSIN II ,GLIOBLASTOMA multiforme ,BRAIN tumors ,RENIN-angiotensin system ,CENTRAL nervous system - Abstract
Glioblastoma (GBM) is an aggressive malignant primary brain tumor with limited therapeutic options. We show that the angiotensin II (AngII) type 2 receptor (AT
2 R) is a therapeutic target for GBM and that AngII, endogenously produced in GBM cells, promotes proliferation through AT2 R. We repurposed EMA401, an AT2 R antagonist originally developed as a peripherally restricted analgesic, for GBM and showed that it inhibits the proliferation of AT2 R-expressing GBM spheroids and blocks their invasiveness and angiogenic capacity. The crystal structure of AT2 R bound to EMA401 was determined and revealed the receptor to be in an active-like conformation with helix-VIII blocking G-protein or ß-arrestin recruitment. The architecture and interactions of EMA401 in AT2 R differ drastically from complexes of AT2 R with other relevant compounds. To enhance central nervous system (CNS) penetration of EMA401, we exploited the crystal structure to design an angiopep-2-tethered EMA401 derivative, A3E. A3E exhibited enhanced CNS penetration, leading to reduced tumor volume, inhibition of proliferation, and increased levels of apoptosis in an orthotopic xenograft model of GBM. [ABSTRACT FROM AUTHOR]- Published
- 2022
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31. Structural basis of the activation of a metabotropic GABA receptor
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Jean-Philippe Pin, Li Xue, Vadim Cherezov, Cornelius Gati, Andrii Ishchenko, Jordy Homing Lam, Philippe Rondard, Gye Won Han, Vsevolod Katritch, Hamidreza Shaye, University of Southern California (USC), Institut de Génomique Fonctionnelle (IGF), Université de Montpellier (UM)-Université Montpellier 1 (UM1)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Université Montpellier 2 - Sciences et Techniques (UM2)-Centre National de la Recherche Scientifique (CNRS), and Stanford University [Stanford]
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Models, Molecular ,0301 basic medicine ,Agonist ,Allosteric modulator ,medicine.drug_class ,Protein subunit ,[SDV]Life Sciences [q-bio] ,Article ,Structure-Activity Relationship ,03 medical and health sciences ,0302 clinical medicine ,Allosteric Regulation ,Protein Domains ,medicine ,Humans ,Receptor ,Binding Sites ,Multidisciplinary ,Chemistry ,Cryoelectron Microscopy ,Transmembrane protein ,Transmembrane domain ,030104 developmental biology ,Metabotropic receptor ,Receptors, GABA-B ,GABA-B Receptor Agonists ,Biophysics ,Protein Multimerization ,Signal transduction ,Apoproteins ,030217 neurology & neurosurgery ,Signal Transduction - Abstract
International audience; Metabotropic γ-aminobutyric acid receptors (GABAB) are involved in the modulation of synaptic responses in the central nervous system and have been implicated in neuropsychological conditions that range from addiction to psychosis1. GABAB belongs to class C of the G-protein-coupled receptors, and its functional entity comprises an obligate heterodimer that is composed of the GB1 and GB2 subunits2. Each subunit possesses an extracellular Venus flytrap domain, which is connected to a canonical seven-transmembrane domain. Here we present four cryo-electron microscopy structures of the human full-length GB1-GB2 heterodimer: one structure of its inactive apo state, two intermediate agonist-bound forms and an active form in which the heterodimer is bound to an agonist and a positive allosteric modulator. The structures reveal substantial differences, which shed light on the complex motions that underlie the unique activation mechanism of GABAB. Our results show that agonist binding leads to the closure of the Venus flytrap domain of GB1, triggering a series of transitions, first rearranging and bringing the two transmembrane domains into close contact along transmembrane helix 6 and ultimately inducing conformational rearrangements in the GB2 transmembrane domain via a lever-like mechanism to initiate downstream signalling. This active state is stabilized by a positive allosteric modulator binding at the transmembrane dimerization interface.
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- 2020
32. Full-length human GLP-1 receptor structure without orthosteric ligands
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Hualiang Jiang, Raymond C. Stevens, Qiansheng Ren, Gye Won Han, Ming-Wei Wang, Nikolaj Kulahin Roed, Steffen Reedtz-Runge, Fan Wu, Michael A. Hanson, Guangyao Lin, Kaini Hang, Gaojie Song, Lijie Wu, Linlin Yang, and Mette Laursen
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0301 basic medicine ,Agonist ,endocrine system ,Protein Conformation ,medicine.drug_class ,G protein ,Science ,General Physics and Astronomy ,Molecular Dynamics Simulation ,Ligands ,Glucagon-Like Peptide-1 Receptor ,Article ,General Biochemistry, Genetics and Molecular Biology ,03 medical and health sciences ,0302 clinical medicine ,Protein structure ,G protein-coupled receptors ,Cryoelectron microscopy ,Receptors, Glucagon ,medicine ,Humans ,Glucose homeostasis ,Amino Acid Sequence ,Disulfides ,lcsh:Science ,Receptor ,Peptide sequence ,Glucagon-like peptide 1 receptor ,X-ray crystallography ,Multidisciplinary ,Protein Stability ,Chemistry ,digestive, oral, and skin physiology ,General Chemistry ,030104 developmental biology ,Membrane protein ,030220 oncology & carcinogenesis ,Biophysics ,lcsh:Q ,Apoproteins ,hormones, hormone substitutes, and hormone antagonists - Abstract
Glucagon-like peptide-1 receptor (GLP-1R) is a class B G protein-coupled receptor that plays an important role in glucose homeostasis and treatment of type 2 diabetes. Structures of full-length class B receptors were determined in complex with their orthosteric agonist peptides, however, little is known about their extracellular domain (ECD) conformations in the absence of orthosteric ligands, which has limited our understanding of their activation mechanism. Here, we report the 3.2 Å resolution, peptide-free crystal structure of the full-length human GLP-1R in an inactive state, which reveals a unique closed conformation of the ECD. Disulfide cross-linking validates the physiological relevance of the closed conformation, while electron microscopy (EM) and molecular dynamic (MD) simulations suggest a large degree of conformational dynamics of ECD that is necessary for binding GLP-1. Our inactive structure represents a snapshot of the peptide-free GLP-1R and provides insights into the activation pathway of this receptor family., Glucagon-like peptide-1 receptor (GLP-1R) plays an important role in glucose homeostasis and treatment of type 2 diabetes. Here authors report the peptide-free crystal structure of human GLP-1R in an inactive state which reveals a unique closed conformation of the extracellular domain.
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- 2020
33. Structural basis of ligand selectivity and disease mutations in cysteinyl leukotriene receptors
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Philippe Sarret, Kirill Kovalev, Alexey Mishin, Toru Maruyama, Aleksandra Luginina, Gye Won Han, Margarita Ergasheva, Egor Marin, Vadim Cherezov, Anastasiia Gusach, Anastasiia Stepko, Nilkanth Patel, Valentin Borshchevskiy, Benjamin Stauch, Élie Besserer-Offroy, Petr Popov, Andrii Ishchenko, Mikhail B. Shevtsov, Taku Fujimoto, Jean-Michel Longpré, Rebecca L. Brouillette, Daria Romanovskaia, Vsevolod Katritch, and Valentin Gordeliy
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Models, Molecular ,0301 basic medicine ,Protein Conformation ,Science ,Central nervous system ,General Physics and Astronomy ,Crystallography, X-Ray ,Ligands ,Protein Engineering ,medicine.disease_cause ,Article ,General Biochemistry, Genetics and Molecular Biology ,Leukotriene D4 ,03 medical and health sciences ,0302 clinical medicine ,Protein structure ,G protein-coupled receptors ,Sf9 Cells ,medicine ,Animals ,Humans ,Computer Simulation ,lcsh:Science ,Receptor ,X-ray crystallography ,Receptors, Leukotriene ,Mutation ,Multidisciplinary ,Chemistry ,Mutagenesis ,HEK 293 cells ,General Chemistry ,Protein engineering ,Asthma ,3. Good health ,Molecular Docking Simulation ,HEK293 Cells ,030104 developmental biology ,medicine.anatomical_structure ,Membrane protein ,Cancer research ,lcsh:Q ,ddc:500 ,030217 neurology & neurosurgery - Abstract
Cysteinyl leukotriene G protein-coupled receptors CysLT1 and CysLT2 regulate pro-inflammatory responses associated with allergic disorders. While selective inhibition of CysLT1R has been used for treating asthma and associated diseases for over two decades, CysLT2R has recently started to emerge as a potential drug target against atopic asthma, brain injury and central nervous system disorders, as well as several types of cancer. Here, we describe four crystal structures of CysLT2R in complex with three dual CysLT1R/CysLT2R antagonists. The reported structures together with the results of comprehensive mutagenesis and computer modeling studies shed light on molecular determinants of CysLTR ligand selectivity and specific effects of disease-related single nucleotide variants., Cysteinyl leukotriene G protein-coupled receptors CysLT1 and CysLT2 regulate pro-inflammatory responses associated with allergic disorders. Here, authors describe four crystal structures of CysLT2R in complex with three dual CysLT1R/CysLT2R antagonists, which shed light on CysLTR ligand selectivity.
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- 2019
34. Crystal structure of the Frizzled 4 receptor in a ligand-free state
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H. Eric Xu, Lu Yang, Petr Popov, Raymond C. Stevens, Fei Xu, Mengchen Pu, Yuanzheng He, Yuxiang Chen, Kelly Suino-Powell, Shaowei Dong, Kaleeckal G. Harikumar, Yiran Wu, Parker W. de Waal, Laurence J. Miller, Zachary J. DeBruine, Yu Guo, Karsten Melcher, Vsevolod Katritch, Wenqing Shui, Gye Won Han, Shifan Yang, Saheem A. Zaidi, Suwen Zhao, Bingjie Zhang, and Ting-Hai Xu
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Models, Molecular ,0301 basic medicine ,Frizzled ,Binding Sites ,Multidisciplinary ,Chemistry ,Protein domain ,Dishevelled Proteins ,Wnt signaling pathway ,Molecular Dynamics Simulation ,Crystallography, X-Ray ,Ligands ,Ligand (biochemistry) ,Frizzled Receptors ,Transmembrane protein ,03 medical and health sciences ,Transmembrane domain ,030104 developmental biology ,Protein Domains ,Biophysics ,Humans ,Cysteine ,Binding site ,Wnt Signaling Pathway ,G protein-coupled receptor - Abstract
Frizzled receptors (FZDs) are class-F G-protein-coupled receptors (GPCRs) that function in Wnt signalling and are essential for developing and adult organisms1,2. As central mediators in this complex signalling pathway, FZDs serve as gatekeeping proteins both for drug intervention and for the development of probes in basic and in therapeutic research. Here we present an atomic-resolution structure of the human Frizzled 4 receptor (FZD4) transmembrane domain in the absence of a bound ligand. The structure reveals an unusual transmembrane architecture in which helix VI is short and tightly packed, and is distinct from all other GPCR structures reported so far. Within this unique transmembrane fold is an extremely narrow and highly hydrophilic pocket that is not amenable to the binding of traditional GPCR ligands. We show that such a pocket is conserved across all FZDs, which may explain the long-standing difficulties in the development of ligands for these receptors. Molecular dynamics simulations on the microsecond timescale and mutational analysis uncovered two coupled, dynamic kinks located at helix VII that are involved in FZD4 activation. The stability of the structure in its ligand-free form, an unfavourable pocket for ligand binding and the two unusual kinks on helix VII suggest that FZDs may have evolved a novel ligand-recognition and activation mechanism that is distinct from that of other GPCRs.
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- 2018
35. Elucidating the active delta-opioid receptor crystal structure with peptide and small-molecule agonists
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Olivier Van der Poorten, Gye Won Han, Brian J. Holleran, Véronique Blais, Tobias Claff, Vadim Cherezov, Louis Gendron, Raymond C. Stevens, Steven Ballet, Michael A. Hanson, Zhi-Jie Liu, Nilkanth Patel, Christa E. Müller, Lijie Wu, Vsevolod Katritch, Philippe Sarret, Kate L. White, Charlotte Martin, Jing Yu, Chemistry, and WE Academic Unit
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Agonist ,medicine.drug_class ,Receptors, Opioid, mu ,Peptide ,Spodoptera ,Pharmacology ,Crystallography, X-Ray ,Biochemistry ,Anxiolytic ,03 medical and health sciences ,0302 clinical medicine ,Protein Domains ,Structural Biology ,Opioid receptor ,Receptors, Opioid, delta ,Sf9 Cells ,medicine ,Animals ,Humans ,Receptor ,Research Articles ,030304 developmental biology ,chemistry.chemical_classification ,0303 health sciences ,Multidisciplinary ,Chemistry ,Chronic pain ,SciAdv r-articles ,respiratory system ,medicine.disease ,Small molecule ,3. Good health ,Molecular Docking Simulation ,Opioid ,Peptides ,human activities ,030217 neurology & neurosurgery ,Research Article ,medicine.drug - Abstract
Crystal structures provide first atomic-level insights into δ-opioid receptor activation by two structurally diverse agonists., Selective activation of the δ-opioid receptor (DOP) has great potential for the treatment of chronic pain, benefitting from ancillary anxiolytic and antidepressant-like effects. Moreover, DOP agonists show reduced adverse effects as compared to μ-opioid receptor (MOP) agonists that are in the spotlight of the current “opioid crisis.” Here, we report the first crystal structures of the DOP in an activated state, in complex with two relevant and structurally diverse agonists: the potent opioid agonist peptide KGCHM07 and the small-molecule agonist DPI-287 at 2.8 and 3.3 Å resolution, respectively. Our study identifies key determinants for agonist recognition, receptor activation, and DOP selectivity, revealing crucial differences between both agonist scaffolds. Our findings provide the first investigation into atomic-scale agonist binding at the DOP, supported by site-directed mutagenesis and pharmacological characterization. These structures will underpin the future structure-based development of DOP agonists for an improved pain treatment with fewer adverse effects.
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- 2019
36. Structural basis for signal recognition and transduction by platelet-activating-factor receptor
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Linlin Yang, Xianping Wang, Raymond C. Stevens, Ye Zhou, Cuiying Yi, Beili Wu, Qiuxiang Tan, Gye Won Han, Hualiang Jiang, Jianfeng Liu, Xuejun Cai Zhang, Qiang Zhao, Xuemei Li, Can Cao, Huaiyu Yang, Anna Qiao, Minmin Lu, Yongfang Zhao, Yiwei Zhou, Chanjuan Xu, Zihe Rao, and Lingli He
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0301 basic medicine ,Cell signaling ,Conformational change ,Indoles ,Protein Conformation ,Platelet Membrane Glycoproteins ,Crystallography, X-Ray ,Ligands ,Receptors, G-Protein-Coupled ,03 medical and health sciences ,Protein structure ,Structural Biology ,Fluorescence Resonance Energy Transfer ,Humans ,Inverse agonist ,Receptor ,Molecular Biology ,G protein-coupled receptor ,Binding Sites ,Chemistry ,Imidazoles ,Hydrogen Bonding ,Molecular Docking Simulation ,Thiazoles ,030104 developmental biology ,Biophysics ,Platelet-activating factor receptor ,Signal transduction ,Platelet Aggregation Inhibitors ,Signal Transduction - Abstract
Platelet-activating-factor receptor (PAFR) responds to platelet-activating factor (PAF), a phospholipid mediator of cell-to-cell communication that exhibits diverse physiological effects. PAFR is considered an important drug target for treating asthma, inflammation and cardiovascular diseases. Here we report crystal structures of human PAFR in complex with the antagonist SR 27417 and the inverse agonist ABT-491 at 2.8-Å and 2.9-Å resolution, respectively. The structures, supported by molecular docking of PAF, provide insights into the signal-recognition mechanisms of PAFR. The PAFR-SR 27417 structure reveals an unusual conformation showing that the intracellular tips of helices II and IV shift outward by 13 Å and 4 Å, respectively, and helix VIII adopts an inward conformation. The PAFR structures, combined with single-molecule FRET and cell-based functional assays, suggest that the conformational change in the helical bundle is ligand dependent and plays a critical role in PAFR activation, thus greatly extending knowledge about signaling by G-protein-coupled receptors.
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- 2018
37. Crystal structures of agonist-bound human cannabinoid receptor CB1
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Jo-Hao Ho, Spyros P. Nikas, Zhi-Jie Liu, Tian Hua, Yiran Wu, Anisha Korde, Haiguang Liu, Kiran Vemuri, Mengchen Pu, Gye Won Han, Kang Ding, Raymond C. Stevens, Alexandros Makriyannis, Shan Jiang, Xuanxuan Li, Laura M. Bohn, Michael A. Hanson, Suwen Zhao, Robert B. Laprairie, and Lu Qu
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0301 basic medicine ,Agonist ,Multidisciplinary ,Cannabinoid receptor ,Stereochemistry ,medicine.drug_class ,Chemistry ,medicine.medical_treatment ,Partial agonist ,Endocannabinoid system ,03 medical and health sciences ,030104 developmental biology ,mental disorders ,Synthetic cannabinoids ,medicine ,Biophysics ,lipids (amino acids, peptides, and proteins) ,Cannabinoid ,Binding site ,Receptor ,medicine.drug - Abstract
Crystal structures of the human cannabinoid receptor 1 (CB1) bound to the agonists AM11542 and AM841 reveal notable structural rearrangements upon receptor activation, and this flexibility may be a common feature among other G-protein-coupled receptors. The human cannabinoid receptor 1 (CB1) is the main target of the plant cannabinoid Δ9-tetrahydrocannbinol (Δ9-THC), the key psychoactive compound in Cannabis sativa. CB1 is activated by endocannabinoids and is a therapeutic target for pain management, epilepsy and obesity, among others, although an active receptor structure is still lacking. Here, Zhi-Jie Liu and colleagues report the crystal structure of CB1 activated by two potent Δ9-THC derivatives, AM11542 and AM841. Both of these agonists have a gem-dimethyl group on their alkyl chain which leads to significant enhancement in their potency and efficacy. Receptor activation involves large-scale structural rearrangements on both extracellular and cytoplasmic sides and a significant reduction in the size of the binding pocket. These conformational changes involve a novel molecular 'twin toggle switch', the synergistic movement of two key residues during activation, which the authors suggest may be common to other G-protein-coupled receptors. The cannabinoid receptor 1 (CB1) is the principal target of the psychoactive constituent of marijuana, the partial agonist Δ9-tetrahydrocannabinol (Δ9-THC)1. Here we report two agonist-bound crystal structures of human CB1 in complex with a tetrahydrocannabinol (AM11542) and a hexahydrocannabinol (AM841) at 2.80 A and 2.95 A resolution, respectively. The two CB1–agonist complexes reveal important conformational changes in the overall structure, relative to the antagonist-bound state2, including a 53% reduction in the volume of the ligand-binding pocket and an increase in the surface area of the G-protein-binding region. In addition, a ‘twin toggle switch’ of Phe2003.36 and Trp3566.48 (superscripts denote Ballesteros–Weinstein numbering3) is experimentally observed and appears to be essential for receptor activation. The structures reveal important insights into the activation mechanism of CB1 and provide a molecular basis for predicting the binding modes of Δ9-THC, and endogenous and synthetic cannabinoids. The plasticity of the binding pocket of CB1 seems to be a common feature among certain class A G-protein-coupled receptors. These findings should inspire the design of chemically diverse ligands with distinct pharmacological properties.
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- 2017
38. Structure of the full-length glucagon class B G-protein-coupled receptor
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Venkatasubramanian Dharmarajan, Anna Qiao, Xiaoai Wu, Guangyao Lin, Raymond G. Sierra, Michael A. Hanson, Patrick R. Griffin, Gye Won Han, Thomas D. Grant, Hui Zhang, Yanhong Wu, Steffen Reedtz-Runge, Linlin Yang, Haonan Zhang, Dehua Yang, Ming-Wei Wang, Wei Liu, Hualiang Jiang, Huaiyu Yang, Garrett Nelson, Xiaoqing Cai, Yuhui Dong, Raymond C. Stevens, Beili Wu, Antao Dai, Vadim Cherezov, Zhi Geng, Uwe Weierstall, Gaojie Song, Chris de Graaf, Limin Ma, Qiang Zhao, Jesper Lau, Medicinal chemistry, and AIMMS
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0301 basic medicine ,Models, Molecular ,Multidisciplinary ,Protein Conformation ,Protein domain ,Biology ,Ligands ,Article ,3. Good health ,Receptors, G-Protein-Coupled ,03 medical and health sciences ,Transmembrane domain ,030104 developmental biology ,0302 clinical medicine ,Biochemistry ,Biophysics ,Glucose homeostasis ,Receptor ,Glucagon receptor ,Glucagon receptor family ,030217 neurology & neurosurgery ,Glucagon-like peptide 1 receptor ,G protein-coupled receptor - Abstract
The human glucagon receptor, GCGR, belongs to the class B G-protein-coupled receptor family and plays a key role in glucose homeostasis and the pathophysiology of type 2 diabetes. Here we report the 3.0 Å crystal structure of full-length GCGR containing both the extracellular domain and transmembrane domain in an inactive conformation. The two domains are connected by a 12-residue segment termed the stalk, which adopts a β-strand conformation, instead of forming an α-helix as observed in the previously solved structure of the GCGR transmembrane domain. The first extracellular loop exhibits a β-hairpin conformation and interacts with the stalk to form a compact β-sheet structure. Hydrogen-deuterium exchange, disulfide crosslinking and molecular dynamics studies suggest that the stalk and the first extracellular loop have critical roles in modulating peptide ligand binding and receptor activation. These insights into the full-length GCGR structure deepen our understanding of the signalling mechanisms of class B G-protein-coupled receptors.
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- 2017
39. Structural basis of ligand recognition and self-activation of orphan GPR52
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Suwen Zhao, Gye Won Han, Xiaohu Wei, Yang Yue, Fei Xu, Mingyue Li, Shimeng Guo, Ming Lei, Yong Chen, Zhipu Luo, Jian Wu, Shaobai Li, Xin Xie, Niandong Wang, Yiran Wu, and Xi Lin
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Agonist ,Models, Molecular ,medicine.drug_class ,Allosteric regulation ,Amino Acid Motifs ,Crystallography, X-Ray ,Ligands ,Receptors, G-Protein-Coupled ,03 medical and health sciences ,0302 clinical medicine ,Allosteric Regulation ,Heterotrimeric G protein ,medicine ,GTP-Binding Protein alpha Subunits, Gs ,Humans ,Amino Acid Sequence ,Binding site ,030304 developmental biology ,G protein-coupled receptor ,0303 health sciences ,Multidisciplinary ,Binding Sites ,Drug discovery ,Chemistry ,Cryoelectron Microscopy ,Ligand (biochemistry) ,Cell biology ,Signal transduction ,Apoproteins ,030217 neurology & neurosurgery ,Allosteric Site - Abstract
GPR52 is a class-A orphan G-protein-coupled receptor that is highly expressed in the brain and represents a promising therapeutic target for the treatment of Huntington's disease and several psychiatric disorders1,2. Pathological malfunction of GPR52 signalling occurs primarily through the heterotrimeric Gs protein2, but it is unclear how GPR52 and Gs couple for signal transduction and whether a native ligand or other activating input is required. Here we present the high-resolution structures of human GPR52 in three states: a ligand-free state, a Gs-coupled self-activation state and a potential allosteric ligand-bound state. Together, our structures reveal that extracellular loop 2 occupies the orthosteric binding pocket and operates as a built-in agonist, conferring an intrinsically high level of basal activity to GPR523. A fully active state is achieved when Gs is coupled to GPR52 in the absence of an external agonist. The receptor also features a side pocket for ligand binding. These insights into the structure and function of GPR52 could improve our understanding of other self-activated GPCRs, enable the identification of endogenous and tool ligands, and guide drug discovery efforts that target GPR52.
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- 2019
40. Structure-based mechanism of cysteinyl leukotriene receptor inhibition by antiasthmatic drugs
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Élie Besserer-Offroy, Egor Marin, Vsevolod Katritch, Valentin Gordeliy, Alexey Mishin, Philippe Sarret, Alexander Batyuk, Vitaly Polovinkin, Petr Popov, Nilkanth Patel, Valentin Borshchevskiy, Nadezhda Safronova, Rebecca L. Brouillette, Uwe Weierstall, Evelina Edelweiss, Vadim Cherezov, Aleksandra Luginina, Anna Shiriaeva, Jean-Michel Longpré, Elizaveta Lyapina, Andrey Bogorodskiy, Wei Liu, Gye Won Han, Hao Hu, Anastasiia Gusach, and Andrii Ishchenko
- Subjects
Indoles ,Phenylcarbamates ,Pharmacology ,Crystallography, X-Ray ,Ligands ,Pranlukast ,Tosyl Compounds ,03 medical and health sciences ,0302 clinical medicine ,Structural Biology ,medicine ,Humans ,Antiasthmatic drugs ,Anti-Asthmatic Agents ,Zafirlukast ,Research Articles ,030304 developmental biology ,Receptors, Leukotriene ,Sulfonamides ,0303 health sciences ,Binding Sites ,Multidisciplinary ,Mechanism (biology) ,Chemistry ,Sodium ,SciAdv r-articles ,Ligand (biochemistry) ,Recombinant Proteins ,Protein Structure, Tertiary ,3. Good health ,Molecular Docking Simulation ,Transmembrane domain ,Membrane protein ,Chromones ,Leukotriene Antagonists ,ddc:500 ,030217 neurology & neurosurgery ,Cysteinyl leukotriene receptor ,Research Article ,medicine.drug - Abstract
Two distinct antagonist-bound structures of CysLT1R reveal unique ligand-binding modes and signaling mechanisms., The G protein–coupled cysteinyl leukotriene receptor CysLT1R mediates inflammatory processes and plays a major role in numerous disorders, including asthma, allergic rhinitis, cardiovascular disease, and cancer. Selective CysLT1R antagonists are widely prescribed as antiasthmatic drugs; however, these drugs demonstrate low effectiveness in some patients and exhibit a variety of side effects. To gain deeper understanding into the functional mechanisms of CysLTRs, we determined the crystal structures of CysLT1R bound to two chemically distinct antagonists, zafirlukast and pranlukast. The structures reveal unique ligand-binding modes and signaling mechanisms, including lateral ligand access to the orthosteric pocket between transmembrane helices TM4 and TM5, an atypical pattern of microswitches, and a distinct four-residue–coordinated sodium site. These results provide important insights and structural templates for rational discovery of safer and more effective drugs.
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- 2019
41. Structural Basis for Ligand Recognition and Functional Selectivity at Angiotensin Receptor
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Raymond C. Stevens, Gye Won Han, Vadim Cherezov, Vsevolod Katritch, Nilkanth Patel, Haitao Zhang, Hamiyet Unal, Sadashiva S. Karnik, and Russell Desnoyer
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Agonist ,Angiotensin receptor ,medicine.drug_class ,Allosteric regulation ,Tetrazoles ,Crystallography, X-Ray ,Ligands ,Binding, Competitive ,Biochemistry ,Partial agonist ,Receptor, Angiotensin, Type 1 ,Cell Line ,Chlorocebus aethiops ,Sf9 Cells ,medicine ,Functional selectivity ,Animals ,Humans ,Inverse agonist ,Computer Simulation ,Molecular Biology ,Antihypertensive Agents ,Ions ,Chemistry ,Cell Membrane ,Sodium ,Imidazoles ,Cell Biology ,Angiotensin II ,Protein Structure, Tertiary ,Protein Structure and Folding ,COS Cells ,Mutation ,Mutagenesis, Site-Directed ,Olmesartan ,Oligopeptides ,Allosteric Site ,Protein Binding ,medicine.drug - Abstract
Angiotensin II type 1 receptor (AT(1)R) is the primary blood pressure regulator. AT(1)R blockers (ARBs) have been widely used in clinical settings as anti-hypertensive drugs and share a similar chemical scaffold, although even minor variations can lead to distinct therapeutic efficacies toward cardiovascular etiologies. The structural basis for AT(1)R modulation by different peptide and non-peptide ligands has remained elusive. Here, we report the crystal structure of the human AT(1)R in complex with an inverse agonist olmesartan (Benicar (TM)), a highly potent anti-hypertensive drug. Olmesartan is anchored to the receptor primarily by the residues Tyr-35(1.39), Trp-84(2.60), and Arg-167(ECL2), similar to the antagonist ZD7155, corroborating a common binding mode of different ARBs. Using docking simulations and site-directed mutagenesis, we identified specific interactions between AT(1)R and different ARBs, including olmesartan derivatives with inverse agonist, neutral antagonist, or agonist activities. We further observed that the mutation N111(3.35)A in the putative sodium-binding site affects binding of the endogenous peptide agonist angiotensin II but not the beta-arrestin-biased peptide TRV120027.
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- 2015
42. An orthogonal seryl-tRNA synthetase/tRNA pair for noncanonical amino acid mutagenesis in Escherichia coli
- Author
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Gye Won Han, Claudio Zambaldo, Peter G. Schultz, Raymond C. Stevens, Fariborz Nasertorabi, Abhishek Chatterjee, and Minseob Koh
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Serine-tRNA Ligase ,Stereochemistry ,Clinical Biochemistry ,Pharmaceutical Science ,Protein Engineering ,medicine.disease_cause ,01 natural sciences ,Biochemistry ,Article ,RNA, Transfer ,SERYL-tRNA SYNTHETASE ,Drug Discovery ,Escherichia coli ,medicine ,Amino Acids ,Molecular Biology ,chemistry.chemical_classification ,Methanosarcina mazei ,biology ,010405 organic chemistry ,Chemistry ,Organic Chemistry ,Mutagenesis ,Archaeoglobus fulgidus ,Active site ,0104 chemical sciences ,Amino acid ,010404 medicinal & biomolecular chemistry ,Methanosarcina ,Transfer RNA ,biology.protein ,Molecular Medicine - Abstract
We report the development of the orthogonal amber-suppressor pair Archaeoglobus fulgidus seryl-tRNA (Af-tRNASer)/Methanosarcina mazei seryl-tRNA synthetase (MmSerRS) in Escherichia coli. Furthermore, the crystal structure of MmSerRS was solved at 1.45 A resolution, which should enable structure-guided engineering of its active site to genetically encode small, polar noncanonical amino acids (ncAAs).
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- 2020
43. Molecular Mechanism for Ligand Recognition and Subtype Selectivity of α2C Adrenergic Receptor
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Lu Qu, Yaping Sun, Houchao Tao, Qianqian Sun, Dong Wu, Suwen Zhao, Xiaoyu Chen, Yunpeng Song, Jie Yang, Lijie Wu, Guisheng Zhong, Shuguang Yuan, Yu Guo, Ling Wang, Qingtong Zhou, Cenfeng Chu, Liu Yang, Zhi-Jie Liu, Raymond C. Stevens, Tao Hu, Gye Won Han, Liaoyuan Hu, Yiran Wu, Yueming Xu, and Quan Wang
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0301 basic medicine ,Cell signaling ,Adrenergic receptor ,Chemistry ,Antagonist ,Adrenergic ,Endogeny ,Ligand (biochemistry) ,General Biochemistry, Genetics and Molecular Biology ,Cell biology ,03 medical and health sciences ,030104 developmental biology ,0302 clinical medicine ,lcsh:Biology (General) ,Receptor ,lcsh:QH301-705.5 ,030217 neurology & neurosurgery ,G protein-coupled receptor - Abstract
Summary: Adrenergic G-protein-coupled receptors (GPCRs) mediate different cellular signaling pathways in the presence of endogenous catecholamines and play important roles in both physiological and pathological conditions. Extensive studies have been carried out to investigate the structure and function of β adrenergic receptors (βARs). However, the structure of α adrenergic receptors (αARs) remains to be determined. Here, we report the structure of the human α2C adrenergic receptor (α2CAR) with the non-selective antagonist, RS79948, at 2.8 Å. Our structure, mutations, modeling, and functional experiments indicate that a α2CAR-specific D206ECL2-R409ECL3-Y4056.58 network plays a role in determining α2 adrenergic subtype selectivity. Furthermore, our results show that a specific loosened helix at the top of TM4 in α2CAR is involved in receptor activation. Together, our structure of human α2CAR-RS79948 provides key insight into the mechanism underlying the α2 adrenergic receptor activation and subtype selectivity. : Chen et al. report the crystal structure of human α2CAR and the functional experimental results, which indicate that extracellular regions determine α2 adrenergic subtype selectivity. The structural and functional results provide the molecular explanation for α2CAR selective ligands and insights to understand GPCR subtype selectivity. Keywords: α2C adrenergic receptor, subtype selectivity, crystal structure, GPCRs, Raynaud's syndrome, JP1302
- Published
- 2019
44. Structural basis of ligand recognition at the human MT
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Benjamin, Stauch, Linda C, Johansson, John D, McCorvy, Nilkanth, Patel, Gye Won, Han, Xi-Ping, Huang, Cornelius, Gati, Alexander, Batyuk, Samuel T, Slocum, Andrii, Ishchenko, Wolfgang, Brehm, Thomas A, White, Nairie, Michaelian, Caleb, Madsen, Lan, Zhu, Thomas D, Grant, Jessica M, Grandner, Anna, Shiriaeva, Reid H J, Olsen, Alexandra R, Tribo, Saïd, Yous, Raymond C, Stevens, Uwe, Weierstall, Vsevolod, Katritch, Bryan L, Roth, Wei, Liu, and Vadim, Cherezov
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Models, Molecular ,Lasers ,Receptor, Melatonin, MT1 ,Electrons ,Ligands ,Antidepressive Agents ,Substrate Specificity ,Molecular Docking Simulation ,Structure-Activity Relationship ,Indenes ,Acetamides ,Mutation ,Receptor, Serotonin, 5-HT2C ,Humans ,Amino Acid Sequence ,Crystallization ,Melatonin - Abstract
Melatonin (N-acetyl-5-methoxytryptamine) is a neurohormone that maintains circadian rhythms
- Published
- 2018
45. Crystal structure of misoprostol bound to the labor inducer prostaglandin E
- Author
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Kate L. White, Vsevolod Katritch, Billy Breton, Martin Audet, Wei Liu, Uwe Weierstall, Wenqing Shui, David Manahan, Barbara Zarzycka, Yan Lu, Raymond C. Stevens, Michael A. Hanson, Vadim Cherezov, Alexander Batyuk, Jeffrey Velasquez, Petr Popov, Cornelius Gati, Gye Won Han, and Hao Hu
- Subjects
Agonist ,medicine.drug_class ,Protein Conformation ,Prostaglandin E2 receptor ,medicine.medical_treatment ,Uterotonic ,Pharmacology ,Crystallography, X-Ray ,Article ,Dinoprostone ,03 medical and health sciences ,Protein structure ,medicine ,Humans ,Inducer ,Receptor ,Molecular Biology ,Misoprostol ,030304 developmental biology ,0303 health sciences ,Binding Sites ,Chemistry ,030302 biochemistry & molecular biology ,Water ,Cell Biology ,3. Good health ,Molecular Docking Simulation ,Receptors, Prostaglandin E, EP3 Subtype ,Mutagenesis, Site-Directed ,Prostaglandin E ,medicine.drug ,Signal Transduction - Abstract
Misoprostol is a life-saving drug in many developing countries for women at risk of post-partum hemorrhaging owing to its affordability, stability, ease of administration and clinical efficacy. However, misoprostol lacks receptor and tissue selectivities, and thus its use is accompanied by a number of serious side effects. The development of pharmacological agents combining the advantages of misoprostol with improved selectivity is hindered by the absence of atomic details of misoprostol action in labor induction. Here, we present the 2.5 A resolution crystal structure of misoprostol free-acid form bound to the myometrium labor-inducing prostaglandin E2 receptor 3 (EP3). The active state structure reveals a completely enclosed binding pocket containing a structured water molecule that coordinates misoprostol's ring structure. Modeling of selective agonists in the EP3 structure reveals rationales for selectivity. These findings will provide the basis for the next generation of uterotonic drugs that will be suitable for administration in low resource settings. A structure of the prostaglandin E2 receptor 3 (EP3) bound to the agonist misoprostol shows a completely enclosed binding pocket with a structured water molecule that coordinates misoprostol's ring structure and explains the receptor's selectivity.
- Published
- 2018
46. Structure of the Nanobody-Stabilized Active State of the Kappa Opioid Receptor
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Gye Won Han, Vsevolod Katritch, Alexandra R. Tribo, F. Ivy Carroll, Vadim Cherezov, Pauline W. Ondachi, Brian E. Krumm, Els Pardon, Sheng Wang, Raymond C. Stevens, Bryan L. Roth, Gavril W. Pasternak, Rajendra Uprety, Eyal Vardy, Ryan T. Strachan, Daniel Wacker, Ming-Yue Lee, Xi Ping Huang, Tao Che, Saheem A. Zaidi, Jan Steyaert, John D. McCorvy, Susruta Majumdar, Philip D. Mosier, Department of Bio-engineering Sciences, and Structural Biology Brussels
- Subjects
0301 basic medicine ,Agonist ,Morphinan ,medicine.drug_class ,Addiction ,Biology ,Molecular Dynamics Simulation ,Spodoptera ,κ-opioid receptor ,General Biochemistry, Genetics and Molecular Biology ,Article ,structure-function ,03 medical and health sciences ,chemistry.chemical_compound ,0302 clinical medicine ,GPCR ,Opioid receptor ,medicine ,Sf9 Cells ,Animals ,Humans ,Receptor ,G protein-coupled receptor ,Analgesics ,Binding Sites ,Crystallography ,Protein Stability ,Biochemistry, Genetics and Molecular Biology(all) ,Receptors, Opioid, kappa ,opioid receptor ,Molecular Docking Simulation ,030104 developmental biology ,HEK293 Cells ,active state ,chemistry ,Morphinans ,Nanobody ,morphinan ,Neuroscience ,030217 neurology & neurosurgery ,Function (biology) ,Intracellular ,Protein Binding - Abstract
The κ-opioid receptor (KOP) mediates the actions of opioids with hallucinogenic, dysphoric, and analgesic activities. The design of KOP analgesics devoid of hallucinatory and dysphoric effects has been hindered by an incomplete structural and mechanistic understanding of KOP agonist actions. Here, we provide a crystal structure of human KOP in complex with the potent epoxymorphinan opioid agonist MP1104 and an active-state-stabilizing nanobody. Comparisons between inactive- and active-state opioid receptor structures reveal substantial conformational changes in the binding pocket and intracellular and extracellular regions. Extensive structural analysis and experimental validation illuminate key residues that propagate larger-scale structural rearrangements and transducer binding that, collectively, elucidate the structural determinants of KOP pharmacology, function, and biased signaling. These molecular insights promise to accelerate the structure-guided design of safer and more effective κ-opioid receptor therapeutics. A crystal structure of the active κ-opioid receptor provides a guide for the development of safe and effective new analgesics. © 2017 Elsevier Inc.
- Published
- 2018
47. Allosteric Coupling of Drug Binding and Intracellular Signaling in the A2A Adenosine Receptor
- Author
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Zhan-Guo Gao, Kyle M. McClary, Kurt Wüthrich, Tatiana Didenko, Gye Won Han, Pawel Stanczak, Raymond C. Stevens, Kenneth A. Jacobson, Martin Audet, Matthew T. Eddy, Ming-Yue Lee, and Reto Horst
- Subjects
Chemistry ,Allosteric regulation ,Biophysics ,Extracellular ,Drug Binding Site ,Adenosine A2A receptor ,Receptor ,Adenosine receptor ,Intracellular ,G protein-coupled receptor - Abstract
Structural plasticity enabling signal transfer over 30 A from the extracellular drug binding site of the adenosine A2A receptor (A2AAR) to the intracellular surface was characterized using NMR spectroscopy in solution. Based on Trp and Gly assignments throughout the receptor, we discovered local structural polymorphisms related to allosteric coupling between the orthosteric drug binding pocket and the intracellular signaling surface. This provides a dynamic structural basis for the function of the allosteric center Asp2.50. Modification of the allosteric center with replacement by Asn2.50 suppressed the structural polymorphisms on the signaling surface without affecting the conformation at the extracellular surface. Our observations suggest A2AAR extracellular and intracellular regions can function as two semi-independent subdomains for the Asn2.50 variant. As this allosteric center is one of the most highly conserved motifs among class A receptors, this implies a common signaling mechanism among many human GPCRs.
- Published
- 2018
48. Advances in Structure Determination of G Protein-Coupled Receptors by SFX
- Author
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Vadim Cherezov, Gye Won Han, Linda C. Johansson, Benjamin Stauch, Alexander Batyuk, and Andrii Ishchenko
- Subjects
0301 basic medicine ,Frizzled ,Chemistry ,Rational design ,SUPERFAMILY ,Computational biology ,03 medical and health sciences ,030104 developmental biology ,0302 clinical medicine ,Structural biology ,Membrane protein ,Signal transduction ,Receptor ,030217 neurology & neurosurgery ,G protein-coupled receptor - Abstract
G protein-coupled receptors (GPCRs) constitute the largest superfamily of membrane proteins, members of which are involved in regulation of critical sensory and physiological processes in the human body. High-resolution GPCR structures are essential for the elucidation of the molecular mechanisms of signal transduction, and for the rational design of more effective therapeutics. GPCR structure determination is, however, hampered by challenges in their expression, stabilization, and crystallization. The recent emergence of X-ray free electron lasers (FELs), and establishment of serial femtosecond crystallography (SFX) have advanced the field of structural biology by enabling access to high-resolution structure and dynamics of challenging to crystallize and radiation damage-sensitive macromolecules. In this chapter we outline relevant SFX technology developments and its applications to structural studies of GPCRs, shedding light on ligand binding to antitumor and anti-addiction targets, uncovering molecular mechanisms behind distinct functions of angiotensin receptors, elucidating full-length structures of multidomain class B and Frizzled receptors, and revealing details of interactions between GPCRs and arrestins.
- Published
- 2018
49. Crystal Structure of Antagonist Bound Human Lysophosphatidic Acid Receptor 1
- Author
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Shinji Nakade, Hirotaka Mizuno, Christopher B. Roth, Raymond C. Stevens, Mauro Mileni, Dora Warshaviak, Guillermo Asmar-Rovira, Caroline Rodgers, Jerold Chun, Michael A. Hanson, Gye Won Han, Rie Omi, Jeffrey Velasquez, Mark T. Griffith, Haruto Kurata, Masahiko Terakado, Yasuyuki Kihara, and Jill E. Chrencik
- Subjects
Models, Molecular ,Cell signaling ,Binding Sites ,Sphingosine ,Ligand ,Biochemistry, Genetics and Molecular Biology(all) ,Biology ,Crystallography, X-Ray ,Ligands ,Endocannabinoid system ,Article ,General Biochemistry, Genetics and Molecular Biology ,Cell biology ,Small Molecule Libraries ,Receptors, Lysosphingolipid ,chemistry.chemical_compound ,chemistry ,Biochemistry ,Lysophosphatidic acid ,Chromatography, Gel ,Humans ,Receptors, Lysophosphatidic Acid ,Binding site ,Autotaxin ,Receptor - Abstract
SummaryLipid biology continues to emerge as an area of significant therapeutic interest, particularly as the result of an enhanced understanding of the wealth of signaling molecules with diverse physiological properties. This growth in knowledge is epitomized by lysophosphatidic acid (LPA), which functions through interactions with at least six cognate G protein-coupled receptors. Herein, we present three crystal structures of LPA1 in complex with antagonist tool compounds selected and designed through structural and stability analyses. Structural analysis combined with molecular dynamics identified a basis for ligand access to the LPA1 binding pocket from the extracellular space contrasting with the proposed access for the sphingosine 1-phosphate receptor. Characteristics of the LPA1 binding pocket raise the possibility of promiscuous ligand recognition of phosphorylated endocannabinoids. Cell-based assays confirmed this hypothesis, linking the distinct receptor systems through metabolically related ligands with potential functional and therapeutic implications for treatment of disease.
- Published
- 2015
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50. Exploring the potential impact of an expanded genetic code on protein function
- Author
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Sean A. Reed, Sei-hyun Choi, Peter G. Schultz, Raymond C. Stevens, Fariborz Nasertorabi, Han Xiao, and Gye Won Han
- Subjects
Models, Molecular ,Protein Conformation ,Genetic Vectors ,Mutant ,Catalysis ,beta-Lactamases ,Evolution, Molecular ,Protein structure ,Transacylation ,X-Ray Diffraction ,Escherichia coli ,Amino Acids ,Cloning, Molecular ,Expanded genetic code ,chemistry.chemical_classification ,Cephalexin ,Multidisciplinary ,Molecular Structure ,biology ,Active site ,Biological Sciences ,Genetic code ,Amino acid ,Kinetics ,chemistry ,Biochemistry ,Genetic Code ,Mutation (genetic algorithm) ,biology.protein ,Electrophoresis, Polyacrylamide Gel ,Crystallization - Abstract
With few exceptions, all living organisms encode the same 20 canonical amino acids; however, it remains an open question whether organisms with additional amino acids beyond the common 20 might have an evolutionary advantage. Here, we begin to test that notion by making a large library of mutant enzymes in which 10 structurally distinct noncanonical amino acids were substituted at single sites randomly throughout TEM-1 β-lactamase. A screen for growth on the β-lactam antibiotic cephalexin afforded a unique p-acrylamido-phenylalanine (AcrF) mutation at Val-216 that leads to an increase in catalytic efficiency by increasing kcat, but not significantly affecting KM. To understand the structural basis for this enhanced activity, we solved the X-ray crystal structures of the ligand-free mutant enzyme and of the deacylation-defective wild-type and mutant cephalexin acyl-enzyme intermediates. These structures show that the Val-216-AcrF mutation leads to conformational changes in key active site residues-both in the free enzyme and upon formation of the acyl-enzyme intermediate-that lower the free energy of activation of the substrate transacylation reaction. The functional changes induced by this mutation could not be reproduced by substitution of any of the 20 canonical amino acids for Val-216, indicating that an expanded genetic code may offer novel solutions to proteins as they evolve new activities.
- Published
- 2015
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