19 results on '"Carlo C. dela Seña"'
Search Results
2. Author Correction: Pharmacological inhibition of PRMT7 links arginine monomethylation to the cellular stress response
- Author
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Magdalena M. Szewczyk, Yoshinori Ishikawa, Shawna Organ, Nozomu Sakai, Fengling Li, Levon Halabelian, Suzanne Ackloo, Amber L. Couzens, Mohammad Eram, David Dilworth, Hideto Fukushi, Rachel Harding, Carlo C. dela Seña, Tsukasa Sugo, Kozo Hayashi, David McLeod, Carlos Zepeda, Ahmed Aman, Maria Sánchez-Osuna, Eric Bonneil, Shinji Takagi, Rima Al-Awar, Mike Tyers, Stephane Richard, Masayuki Takizawa, Anne-Claude Gingras, Cheryl H. Arrowsmith, Masoud Vedadi, Peter J. Brown, Hiroshi Nara, and Dalia Barsyte-Lovejoy
- Subjects
Science - Abstract
An amendment to this paper has been published and can be accessed via a link at the top of the paper.
- Published
- 2020
- Full Text
- View/download PDF
3. Correction: A chemical probe of CARM1 alters epigenetic plasticity against breast cancer cell invasion
- Author
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Xiao-Chuan Cai, Tuo Zhang, Eui-jun Kim, Ming Jiang, Ke Wang, Junyi Wang, Shi Chen, Nawei Zhang, Hong Wu, Fengling Li, Carlo C dela Seña, Hong Zeng, Victor Vivcharuk, Xiang Niu, Weihong Zheng, Jonghan P Lee, Yuling Chen, Dalia Barsyte, Magda Szewczyk, Taraneh Hajian, Glorymar Ibáñez, Aiping Dong, Ludmila Dombrovski, Zhenyu Zhang, Haiteng Deng, Jinrong Min, Cheryl H Arrowsmith, Linas Mazutis, Lei Shi, Masoud Vedadi, Peter J Brown, Jenny Xiang, Li-Xuan Qin, Wei Xu, and Minkui Luo
- Subjects
Medicine ,Science ,Biology (General) ,QH301-705.5 - Published
- 2020
- Full Text
- View/download PDF
4. A chemical probe of CARM1 alters epigenetic plasticity against breast cancer cell invasion
- Author
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Xiao-Chuan Cai, Tuo Zhang, Eui-jun Kim, Ming Jiang, Ke Wang, Junyi Wang, Shi Chen, Nawei Zhang, Hong Wu, Fengling Li, Carlo C dela Seña, Hong Zeng, Victor Vivcharuk, Xiang Niu, Weihong Zheng, Jonghan P Lee, Yuling Chen, Dalia Barsyte, Magda Szewczyk, Taraneh Hajian, Glorymar Ibáñez, Aiping Dong, Ludmila Dombrovski, Zhenyu Zhang, Haiteng Deng, Jinrong Min, Cheryl H Arrowsmith, Linas Mazutis, Lei Shi, Masoud Vedadi, Peter J Brown, Jenny Xiang, Li-Xuan Qin, Wei Xu, and Minkui Luo
- Subjects
PRMT ,methylation ,epigenetic ,inhibitor ,single cell ,mechanism ,Medicine ,Science ,Biology (General) ,QH301-705.5 - Abstract
CARM1 is a cancer-relevant protein arginine methyltransferase that regulates many aspects of transcription. Its pharmacological inhibition is a promising anti-cancer strategy. Here SKI-73 (6a in this work) is presented as a CARM1 chemical probe with pro-drug properties. SKI-73 (6a) can rapidly penetrate cell membranes and then be processed into active inhibitors, which are retained intracellularly with 10-fold enrichment for several days. These compounds were characterized for their potency, selectivity, modes of action, and on-target engagement. SKI-73 (6a) recapitulates the effect of CARM1 knockout against breast cancer cell invasion. Single-cell RNA-seq analysis revealed that the SKI-73(6a)-associated reduction of invasiveness acts by altering epigenetic plasticity and suppressing the invasion-prone subpopulation. Interestingly, SKI-73 (6a) and CARM1 knockout alter the epigenetic plasticity with remarkable difference, suggesting distinct modes of action for small-molecule and genetic perturbations. We therefore discovered a CARM1-addiction mechanism of cancer metastasis and developed a chemical probe to target this process.
- Published
- 2019
- Full Text
- View/download PDF
5. A First-in-Class, Highly Selective and Cell-Active Allosteric Inhibitor of Protein Arginine Methyltransferase 6
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Jian Jin, Levon Halabelian, Fengling Li, Masoud Vedadi, Rima Al-awar, Carlo C. dela Seña, Mohammad S. Eram, Aiping Dong, Carlos Zepeda-Velázquez, He Chen, Matthieu Schapira, Robert M. Campbell, Dalia Barsyte-Lovejoy, David McLeod, Cheryl H. Arrowsmith, Hong Zeng, Hong Wu, Yudao Shen, Mary M. Mader, Viacheslav V. Trush, Brian Morgan Watson, Kwang-Su Park, Irene Chau, Fanye Meng, Peter Brown, H. Ümit Kaniskan, and Magdalena M. Szewczyk
- Subjects
Protein-Arginine N-Methyltransferases ,Methyltransferase ,Arginine ,Stereochemistry ,Cell ,Allosteric regulation ,Crystallography, X-Ray ,01 natural sciences ,Article ,03 medical and health sciences ,Allosteric Regulation ,Drug Discovery ,medicine ,Humans ,Enzyme Inhibitors ,IC50 ,030304 developmental biology ,Benzodiazepinones ,0303 health sciences ,Chemistry ,Nuclear Proteins ,Stereoisomerism ,Highly selective ,0104 chemical sciences ,010404 medicinal & biomolecular chemistry ,HEK293 Cells ,medicine.anatomical_structure ,Molecular Medicine ,Enantiomer ,Selectivity ,Allosteric Site ,Protein Binding - Abstract
Protein arginine methyltransferase 6 (PRMT6) catalyzes monomethylation and asymmetric dimethylation of arginine residues in various proteins, plays important roles in biological processes, and is associated with multiple cancers. To date, a highly selective PRMT6 inhibitor has not been reported. Here we report the discovery and characterization of a first-in-class, highly selective allosteric inhibitor of PRMT6, (R)-2 (SGC6870). (R)-2 is a potent PRMT6 inhibitor (IC(50) = 77 ± 6 nM) with outstanding selectivity for PRMT6 over a broad panel of other methyltransferases and nonepigenetic targets. Notably, the crystal structure of the PRMT6−(R)-2 complex and kinetic studies revealed (R)-2 binds a unique, induced allosteric pocket. Additionally, (R)-2 engages PRMT6 and potently inhibits its methyltransferase activity in cells. Moreover, (R)-2’s enantiomer, (S)-2 (SGC6870N), is inactive against PRMT6 and can be utilized as a negative control. Collectively, (R)-2 is a well-characterized PRMT6 chemical probe and a valuable tool for further investigating PRMT6 functions in health and disease.
- Published
- 2021
6. Pharmacological inhibition of PRMT7 links arginine monomethylation to the cellular stress response
- Author
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Hideto Fukushi, Masayuki Takizawa, Shawna Organ, Masoud Vedadi, Peter Brown, Rima Al-awar, Carlo C. dela Seña, Mike Tyers, Carlos Zepeda, Levon Halabelian, David Dilworth, Suzanne Ackloo, Ahmed Aman, Amber L. Couzens, Yoshinori Ishikawa, David Macleod, Nozomu Sakai, Rachel Harding, Stéphane Richard, Tsukasa Sugo, Anne-Claude Gingras, Hiroshi Nara, Fengling Li, Eric Bonneil, Mohammad S. Eram, Cheryl H. Arrowsmith, Shinji Takagi, Dalia Barsyte-Lovejoy, María Sánchez-Osuna, Kozo Hayashi, and Magdalena M. Szewczyk
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0301 basic medicine ,Methyltransferase ,Arginine ,Science ,Cell ,General Physics and Astronomy ,Methylation ,General Biochemistry, Genetics and Molecular Biology ,Article ,Target validation ,03 medical and health sciences ,Stress signalling ,0302 clinical medicine ,Transferases ,Cellular stress response ,medicine ,lcsh:Science ,Multidisciplinary ,Chemistry ,General Chemistry ,Hsp70 ,Cell biology ,030104 developmental biology ,Proteostasis ,medicine.anatomical_structure ,Proteasome ,030220 oncology & carcinogenesis ,lcsh:Q - Abstract
Protein arginine methyltransferases (PRMTs) regulate diverse biological processes and are increasingly being recognized for their potential as drug targets. Here we report the discovery of a potent, selective, and cell-active chemical probe for PRMT7. SGC3027 is a cell permeable prodrug, which in cells is converted to SGC8158, a potent, SAM-competitive PRMT7 inhibitor. Inhibition or knockout of cellular PRMT7 results in drastically reduced levels of arginine monomethylated HSP70 family stress-associated proteins. Structural and biochemical analyses reveal that PRMT7-driven in vitro methylation of HSP70 at R469 requires an ATP-bound, open conformation of HSP70. In cells, SGC3027 inhibits methylation of both constitutive and inducible forms of HSP70, and leads to decreased tolerance for perturbations of proteostasis including heat shock and proteasome inhibitors. These results demonstrate a role for PRMT7 and arginine methylation in stress response., Protein arginine methyltransferases (PRMTs) are increasingly recognized as potential therapeutic targets but PRMT7 remains an understudied member of this enzyme family. Here, the authors develop a chemical probe for PRMT7 and apply it to elucidate the role of PRMT7 in the cellular stress response.
- Published
- 2020
7. Author Correction: Pharmacological inhibition of PRMT7 links arginine monomethylation to the cellular stress response
- Author
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Levon Halabelian, Nozomu Sakai, David McLeod, Anne-Claude Gingras, Rima Al-awar, Hiroshi Nara, Dalia Barsyte-Lovejoy, Masoud Vedadi, David Dilworth, Hideto Fukushi, Stéphane Richard, Carlos Zepeda, Cheryl H. Arrowsmith, María Sánchez-Osuna, Ahmed Aman, Carlo C. dela Seña, Kozo Hayashi, Suzanne Ackloo, Magdalena M. Szewczyk, Yoshinori Ishikawa, Peter Brown, Fengling Li, Eric Bonneil, Mike Tyers, Mohammad S. Eram, Masayuki Takizawa, Amber L. Couzens, Shawna Organ, Rachel Harding, Shinji Takagi, and Tsukasa Sugo
- Subjects
Protein-Arginine N-Methyltransferases ,Arginine ,Science ,General Physics and Astronomy ,Pharmacology ,Biology ,Methylation ,General Biochemistry, Genetics and Molecular Biology ,Target validation ,Stress signalling ,Text mining ,Transferases ,Stress, Physiological ,Cellular stress response ,Sf9 Cells ,Animals ,Humans ,HSP70 Heat-Shock Proteins ,lcsh:Science ,Author Correction ,Multidisciplinary ,business.industry ,General Chemistry ,HCT116 Cells ,Recombinant Proteins ,Gene Knockdown Techniques ,lcsh:Q ,business ,Protein Processing, Post-Translational - Abstract
Protein arginine methyltransferases (PRMTs) regulate diverse biological processes and are increasingly being recognized for their potential as drug targets. Here we report the discovery of a potent, selective, and cell-active chemical probe for PRMT7. SGC3027 is a cell permeable prodrug, which in cells is converted to SGC8158, a potent, SAM-competitive PRMT7 inhibitor. Inhibition or knockout of cellular PRMT7 results in drastically reduced levels of arginine monomethylated HSP70 family stress-associated proteins. Structural and biochemical analyses reveal that PRMT7-driven in vitro methylation of HSP70 at R469 requires an ATP-bound, open conformation of HSP70. In cells, SGC3027 inhibits methylation of both constitutive and inducible forms of HSP70, and leads to decreased tolerance for perturbations of proteostasis including heat shock and proteasome inhibitors. These results demonstrate a role for PRMT7 and arginine methylation in stress response.
- Published
- 2020
8. TP-064, a potent and selective small molecule inhibitor of PRMT4 for multiple myeloma
- Author
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Dalia Barsyte-Lovejoy, Yuji Baba, Peter Brown, Atsushi Kiba, Daisuke Tomita, Douglas R. Cary, Aiping Dong, Hong Zeng, Mihoko Kunitomo, Kazuhide Nakayama, Cheryl H. Arrowsmith, Fengling Li, Matthieu Schapira, Mohammad S. Eram, Charles E. Grimshaw, Yasuhiro Imaeda, Carlo C. dela Seña, Kumar Singh Saikatendu, Hong Wu, Akihiro Ohashi, Michiko Tawada, Renato Ferreira de Freitas, Magdalena M. Szewczyk, and Masoud Vedadi
- Subjects
0301 basic medicine ,crystal structure ,Methyltransferase ,Arginine ,CARM1 ,Chemistry ,Protein subunit ,small molecule inhibitor ,Cell cycle ,TP-064 ,Small molecule ,3. Good health ,multiple myeloma ,03 medical and health sciences ,030104 developmental biology ,0302 clinical medicine ,Oncology ,Biochemistry ,Cell culture ,030220 oncology & carcinogenesis ,PRMT4 ,IC50 ,Research Paper - Abstract
Protein arginine methyltransferase (PRMT) 4 (also known as coactivator-associated arginine methyltransferase 1; CARM1) is involved in a variety of biological processes and is considered as a candidate oncogene owing to its overexpression in several types of cancer. Selective PRMT4 inhibitors are useful tools for clarifying the molecular events regulated by PRMT4 and for validating PRMT4 as a therapeutic target. Here, we report the discovery of TP-064, a potent, selective, and cell-active chemical probe of human PRMT4 and its co-crystal structure with PRMT4. TP-064 inhibited the methyltransferase activity of PRMT4 with high potency (half-maximal inhibitory concentration, IC50 < 10 nM) and selectivity over other PRMT family proteins, and reduced arginine dimethylation of the PRMT4 substrates BRG1-associated factor 155 (BAF155; IC50= 340 ± 30 nM) and Mediator complex subunit 12 (MED12; IC50 = 43 ± 10 nM). TP-064 treatment inhibited the proliferation of a subset of multiple myeloma cell lines, with affected cells arrested in G1 phase of the cell cycle. TP-064 and its negative control (TP-064N) will be valuable tools to further investigate the biology of PRMT4 and the therapeutic potential of PRMT4 inhibition.
- Published
- 2018
9. A chemical probe of CARM1 alters epigenetic plasticity against breast cancer cell invasion
- Author
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Yuling Chen, Hong Zeng, Xiao-Chuan Cai, Magda Szewczyk, Ludmila Dombrovsky, Linas Mazutis, Ming Jiang, Aiping Dong, Taraneh Hajian, Glorymar Ibáñez, Hong Wu, Peter Brown, Fengling Li, Cheryl H. Arrowsmith, Lei Shi, Masoud Vedadi, Victor Vivcharuk, Jenny Xiang, Haiteng Deng, Nawei Zhang, Carlo C. dela Seña, Tuo Zhang, Li-Xuan Qin, Jinrong Min, Zhenyu Zhang, Dalia Barsyte, Wei Xu, Junyi Wang, Jonghan P Lee, Ke Wang, Weihong Zheng, Shi Chen, Eui-jun Kim, Minkui Luo, and Xiang Niu
- Subjects
0301 basic medicine ,genetic structures ,CARM1 ,QH301-705.5 ,Science ,PRMT ,Cell ,Chemical biology ,Cancer metastasis ,mechanism ,Chemical probe ,Plasticity ,behavioral disciplines and activities ,General Biochemistry, Genetics and Molecular Biology ,03 medical and health sciences ,0302 clinical medicine ,Biochemistry and Chemical Biology ,Transcription (biology) ,medicine ,Epigenetics ,Biology (General) ,030304 developmental biology ,0303 health sciences ,General Immunology and Microbiology ,Chemistry ,General Neuroscience ,General Medicine ,Methylation ,single cell ,3. Good health ,Cell biology ,inhibitor ,030104 developmental biology ,Membrane ,medicine.anatomical_structure ,nervous system ,030220 oncology & carcinogenesis ,Medicine ,Breast cancer cells ,methylation ,human activities ,epigenetic ,psychological phenomena and processes ,Research Article ,Human - Abstract
CARM1 is a cancer-relevant protein arginine methyltransferase that regulates many aspects of transcription. Its pharmacological inhibition is a promising anti-cancer strategy. Here SKI-73 (6a in this work) is presented as a CARM1 chemical probe with pro-drug properties. SKI-73 (6a) can rapidly penetrate cell membranes and then be processed into active inhibitors, which are retained intracellularly with 10-fold enrichment for several days. These compounds were characterized for their potency, selectivity, modes of action, and on-target engagement. SKI-73 (6a) recapitulates the effect of CARM1 knockout against breast cancer cell invasion. Single-cell RNA-seq analysis revealed that the SKI-73(6a)-associated reduction of invasiveness acts by altering epigenetic plasticity and suppressing the invasion-prone subpopulation. Interestingly, SKI-73 (6a) and CARM1 knockout alter the epigenetic plasticity with remarkable difference, suggesting distinct modes of action for small-molecule and genetic perturbations. We therefore discovered a CARM1-addiction mechanism of cancer metastasis and developed a chemical probe to target this process., eLife digest Drugs that are small molecules have the potential to block the individual proteins that drive the spread of cancer, but their design is a challenge. This is because they need to get inside the cell and find their target without binding to other proteins on the way. However, small molecule drugs often have an electric charge, which makes it hard for them to cross the cell membrane. Additionally, most proteins are not completely unique, making it harder for the drugs to find the correct target. CARM1 is a protein that plays a role in the spread of breast cancer cells, and scientists are currently looking for a small molecule that will inhibit its action. The group of enzymes that CARM1 belongs to act by taking a small chemical group, called a methyl group, from a molecule called SAM, and transferring it to proteins that switch genes on and off. In the case of CARM1, this changes cell behavior by turning on genes involved in cell movement. Genetically modifying cells so they will not produce any CARM1 stops the spread of breast cancer cells, but developing a drug with the same effects has proved difficult. Existing drugs that can inhibit CARM1 in a test tube struggle to get inside cells and to distinguish between CARM1 and its related enzymes. Now, Cai et al. have modified and tested a CARM1 inhibitor to address these problems, and find out how these small molecules work. At its core, the inhibitor has a structure very similar to a SAM molecule, so it can fit into the SAM binding pocket of CARM1 and its related enzymes. To stop the inhibitor from binding to other proteins, Cai et al. made small changes to its structure until it only interacted with CARM1.Then, to get the inhibitor inside breast cancer cells, Cai et al. cloaked its charged area with a chemical shield, allowing it to cross the cell membrane. Inside the cell, the chemical shield broke away, allowing the inhibitor to attach to CARM1. Analysis of cells showed that this inhibition only affected the cancer cells most likely to spread. Blocking CARM1 switched off genes involved in cell movement and stopped cancer cells from travelling through 3D gels. This work is a step towards making a drug that can block CARM1 in cancer cells, but there is still further work to be done. The next stages will be to test whether the new inhibitor works in other types of cancer cells, in living animals, and in human patient samples.
- Published
- 2019
10. Author response: A chemical probe of CARM1 alters epigenetic plasticity against breast cancer cell invasion
- Author
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Xiao-Chuan Cai, Tuo Zhang, Eui-jun Kim, Ming Jiang, Ke Wang, Junyi Wang, Shi Chen, Nawei Zhang, Hong Wu, Fengling Li, Carlo C dela Seña, Hong Zeng, Victor Vivcharuk, Xiang Niu, Weihong Zheng, Jonghan P Lee, Yuling Chen, Dalia Barsyte, Magda Szewczyk, Taraneh Hajian, Glorymar Ibáñez, Aiping Dong, Ludmila Dombrovski, Zhenyu Zhang, Haiteng Deng, Jinrong Min, Cheryl H Arrowsmith, Linas Mazutis, Lei Shi, Masoud Vedadi, Peter J Brown, Jenny Xiang, Li-Xuan Qin, Wei Xu, and Minkui Luo
- Published
- 2019
11. Chemical probes for protein arginine methyltransferases
- Author
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Masoud Vedadi, Carlo C. dela Seña, Fengling Li, Alice Shi Ming Li, Mohammad S. Eram, and Albina Bolotokova
- Subjects
Protein-Arginine N-Methyltransferases ,S-Adenosylmethionine ,Methyltransferase ,Arginine ,Chemical probe ,Peptide ,General Biochemistry, Genetics and Molecular Biology ,Histones ,03 medical and health sciences ,Transcription (biology) ,Neoplasms ,Humans ,Enzyme Inhibitors ,Molecular Biology ,Phylogeny ,030304 developmental biology ,Cell Proliferation ,Enzyme Assays ,chemistry.chemical_classification ,0303 health sciences ,Chemistry ,F-Box Proteins ,030302 biochemistry & molecular biology ,DNA Damage Repair ,Kinetics ,Biochemistry ,RNA Splicing Factors ,Methyl donor - Abstract
Protein arginine methyltransferases (PRMTs) catalyze the transfer of methyl groups to specific arginine residues of their substrates using S-adenosylmethionine as a methyl donor, contributing to regulation of many biological processes including transcription, and DNA damage repair. Dysregulation of PRMT expression is often associated with various diseases including cancers. Different methods have been used to characterize the activities of PRMTs and determine their kinetic parameters including mass spectrometry, radiometric, and antibody-based assays. Here, we present kinetic characterization of PRMTs using a radioactivity-based assay for better comparison along with previously reported values. We also report on full characterization of PRMT9 activity with SAP145 peptide as substrate. We further review the potent, selective and cell-active PRMT inhibitors discovered in recent years to provide a better understanding of available tools to investigate the roles these proteins play in health and disease.
- Published
- 2019
12. Identification and characterization of the first fragment hits for SETDB1 Tudor domain
- Author
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Steven Kennedy, Sean K. Liew, Matthieu Schapira, Masoud Vedadi, Abdellah Allali-Hassani, Cheryl H. Arrowsmith, Scott Houliston, Andrei K. Yudin, Jinrong Min, Diego B. Diaz, P. Mader, Hong Wu, Aman Iqbal, Aiping Dong, Vijayaratnam Santhakumar, Rodrigo Mendoza-Sanchez, Renato Ferreira de Freitas, Victoria B. Corless, David Smil, Peter Brown, Ludmila Dombrovski, Elena Dobrovetsky, and Carlo C. dela Seña
- Subjects
Models, Molecular ,Tudor domain ,Methyltransferase ,Clinical Biochemistry ,Pharmaceutical Science ,Peptide binding ,Crystallography, X-Ray ,01 natural sciences ,Biochemistry ,Histones ,Small Molecule Libraries ,03 medical and health sciences ,Structure-Activity Relationship ,0302 clinical medicine ,Drug Discovery ,Transferase ,Humans ,Enzyme Inhibitors ,Molecular Biology ,030304 developmental biology ,0303 health sciences ,biology ,Dose-Response Relationship, Drug ,Molecular Structure ,Tudor Domain ,010405 organic chemistry ,Chemistry ,Organic Chemistry ,Nuclear magnetic resonance spectroscopy ,Histone-Lysine N-Methyltransferase ,Small molecule ,0104 chemical sciences ,3. Good health ,Chromatin ,010404 medicinal & biomolecular chemistry ,Histone ,Acetylation ,030220 oncology & carcinogenesis ,biology.protein ,Molecular Medicine - Abstract
SET domain bifurcated protein 1 (SETDB1) is a human histone-lysine methyltransferase, which is amplified in human cancers and was shown to be crucial in the growth of non-small and small cell lung carcinoma. In addition to its catalytic domain, SETDB1 harbors a unique tandem tudor domain which recognizes histone sequences containing both methylated and acetylated lysines, and likely contributes to its localization on chromatin. Using X-ray crystallography and NMR spectroscopy fragment screening approaches, we have identified the first small molecule fragment hits that bind to histone peptide binding groove of the TTD of SETDB1. Herein, we describe the binding modes of these fragments and analogues and the biophysical characterization of key compounds. These confirmed small molecule fragments will inform the development of potent antagonists of SETDB1 interaction with histones.
- Published
- 2019
13. Pharmacological inhibition of PRMT7 links arginine monomethylation to the cellular stress responses
- Author
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Stéphane Richard, Cheryl H. Arrowsmith, Hiroshi Nara, Carlo C. dela Seña, Shinji Takagi, Carlos Zepeda, Rima Al-awar, Yoshinori Ishikawa, Dalia Barsyte-Lovejoy, David Macleod, Shawna Organ, Suzanne Ackloo, Peter Brown, Fengling Li, Tsukasa Sugo, Masoud Vedadi, Nozomu Sakai, David Dilworth, Mohammad S. Eram, Hideto Fukushi, Kozo Hayashi, Magdalena M. Szewczyk, Masayuki Takizawa, and Rachel Harding
- Subjects
0303 health sciences ,Methyltransferase ,Arginine ,Chemistry ,Cell ,Methylation ,3. Good health ,Hsp70 ,Cell biology ,03 medical and health sciences ,0302 clinical medicine ,Proteostasis ,medicine.anatomical_structure ,Proteasome ,030220 oncology & carcinogenesis ,Cellular stress response ,medicine ,030304 developmental biology - Abstract
Protein arginine methyltransferases (PRMTs) regulate diverse biological processes and are increasingly being recognized for their potential as drug targets. Here we report the discovery of a potent, selective and cell active chemical probe for PRMT7. SGC3027 is a cell permeable prodrug, which in cells, is converted to SGC8158, a potent, SAM-competitive PRMT7 inhibitor. Inhibition or knockout of cellular PRMT7 resulted in drastically reduced levels of arginine monomethylation of HSP70 family members and other stress-associated proteins. Structural and biochemical analysis revealed that PRMT7-driven in vitro methylation of HSP70 at R469 requires an ATP-bound, open conformation of HSP70. In cells, SGC3027 inhibited methylation of both constitutive and inducible forms of HSP70, and led to decreased tolerance for perturbations of proteostasis including heat shock and proteasome inhibitors. These results demonstrate a role for PRMT7 and arginine methylation in stress response.
- Published
- 2018
- Full Text
- View/download PDF
14. A Potent, Selective, and Cell-Active Inhibitor of Human Type I Protein Arginine Methyltransferases
- Author
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Brandon A. Speed, Fengling Li, Mohammad S. Eram, Jing Liu, Kyle V. Butler, Magdalena M. Szewczyk, H. Uemit Kaniskan, Hong Zeng, Peter Brown, Jian Jin, Matthieu Schapira, Hong Wu, Masoud Vedadi, Aiping Dong, Dalia Barsyte-Lovejoy, Guillermo Senisterra, Carlo C. dela Seña, Cheryl H. Arrowsmith, and Yudao Shen
- Subjects
Models, Molecular ,0301 basic medicine ,Protein-Arginine N-Methyltransferases ,Methyltransferase ,Arginine ,Cell ,Antineoplastic Agents ,Human type ,Biology ,Biochemistry ,Article ,03 medical and health sciences ,Cell Line, Tumor ,medicine ,Humans ,Transferase ,Pyrroles ,Cell Proliferation ,Molecular Structure ,Extramural ,General Medicine ,Ethylenediamines ,3. Good health ,030104 developmental biology ,medicine.anatomical_structure ,Ethanolamines ,Cell culture ,Molecular Medicine - Abstract
Protein arginine methyltransferases (PRMTs) play a crucial role in a variety of biological processes. Overexpression of PRMTs has been implicated in various human diseases including cancer. Consequently, selective small-molecule inhibitors of PRMTs have been pursued by both academia and the pharmaceutical industry as chemical tools for testing biological and therapeutic hypotheses. PRMTs are divided into three categories: type I PRMTs which catalyze mono- and asymmetric dimethylation of arginine residues, type II PRMTs which catalyze mono- and symmetric dimethylation of arginine residues, and type III PRMT which catalyzes only monomethylation of arginine residues. Here, we report the discovery of a potent, selective, and cell-active inhibitor of human type I PRMTs, MS023, and characterization of this inhibitor in a battery of biochemical, biophysical, and cellular assays. MS023 displayed high potency for type I PRMTs including PRMT1, -3, -4, -6, and -8 but was completely inactive against type II and type III PRMTs, protein lysine methyltransferases and DNA methyltransferases. A crystal structure of PRMT6 in complex with MS023 revealed that MS023 binds the substrate binding site. MS023 potently decreased cellular levels of histone arginine asymmetric dimethylation. It also reduced global levels of arginine asymmetric dimethylation and concurrently increased levels of arginine monomethylation and symmetric dimethylation in cells. We also developed MS094, a close analog of MS023, which was inactive in biochemical and cellular assays, as a negative control for chemical biology studies. MS023 and MS094 are useful chemical tools for investigating the role of type I PRMTs in health and disease.
- Published
- 2015
15. A First-in-Class, Highly Selective and Cell-Active Allosteric Inhibitor of Protein Arginine Methyltransferase 6.
- Author
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Shen Y, Li F, Szewczyk MM, Halabelian L, Chau I, Eram MS, Dela Seña C, Park KS, Meng F, Chen H, Zeng H, Dong A, Wu H, Trush VV, McLeod D, Zepeda-Velázquez CA, Campbell RM, Mader MM, Watson BM, Schapira M, Arrowsmith CH, Al-Awar R, Barsyte-Lovejoy D, Kaniskan HÜ, Brown PJ, Vedadi M, and Jin J
- Subjects
- Allosteric Regulation, Allosteric Site, Benzodiazepinones chemical synthesis, Benzodiazepinones metabolism, Crystallography, X-Ray, Enzyme Inhibitors chemical synthesis, Enzyme Inhibitors metabolism, HEK293 Cells, Humans, Nuclear Proteins metabolism, Protein Binding, Protein-Arginine N-Methyltransferases metabolism, Stereoisomerism, Benzodiazepinones pharmacology, Enzyme Inhibitors pharmacology, Nuclear Proteins antagonists & inhibitors, Protein-Arginine N-Methyltransferases antagonists & inhibitors
- Abstract
Protein arginine methyltransferase 6 (PRMT6) catalyzes monomethylation and asymmetric dimethylation of arginine residues in various proteins, plays important roles in biological processes, and is associated with multiple cancers. To date, a highly selective PRMT6 inhibitor has not been reported. Here we report the discovery and characterization of a first-in-class, highly selective allosteric inhibitor of PRMT6, (R)- 2 (SGC6870). (R)- 2 is a potent PRMT6 inhibitor (IC
50 = 77 ± 6 nM) with outstanding selectivity for PRMT6 over a broad panel of other methyltransferases and nonepigenetic targets. Notably, the crystal structure of the PRMT6- (R)- 2 complex and kinetic studies revealed (R)- 2 binds a unique, induced allosteric pocket. Additionally, (R)- 2 engages PRMT6 and potently inhibits its methyltransferase activity in cells. Moreover, (R)- 2 's enantiomer, (S)- 2 (SGC6870N), is inactive against PRMT6 and can be utilized as a negative control. Collectively, (R) - 2 is a well-characterized PRMT6 chemical probe and a valuable tool for further investigating PRMT6 functions in health and disease.- Published
- 2021
- Full Text
- View/download PDF
16. Substrate Specificity of Purified Recombinant Chicken β-Carotene 9',10'-Oxygenase (BCO2).
- Author
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Dela Seña C, Sun J, Narayanasamy S, Riedl KM, Yuan Y, Curley RW Jr, Schwartz SJ, and Harrison EH
- Subjects
- Amino Acid Sequence, Animals, Carotenoids chemistry, Carotenoids metabolism, Chickens genetics, Cryptoxanthins chemistry, Cryptoxanthins metabolism, Dioxygenases chemistry, Dioxygenases genetics, Humans, Recombinant Proteins chemistry, Recombinant Proteins genetics, Recombinant Proteins metabolism, Sequence Alignment, Substrate Specificity, beta Carotene chemistry, Chickens metabolism, Dioxygenases metabolism, beta Carotene metabolism
- Abstract
Provitamin A carotenoids are oxidatively cleaved by β-carotene 15,15'-dioxygenase (BCO1) at the central 15-15' double bond to form retinal (vitamin A aldehyde). Another carotenoid oxygenase, β-carotene 9',10'-oxygenase (BCO2) catalyzes the oxidative cleavage of carotenoids at the 9'-10' bond to yield an ionone and an apo-10'-carotenoid. Previously published substrate specificity studies of BCO2 were conducted using crude lysates from bacteria or insect cells expressing recombinant BCO2. Our attempts to obtain active recombinant human BCO2 expressed in Escherichia coli were unsuccessful. We have expressed recombinant chicken BCO2 in the strain E. coli BL21-Gold (DE3) and purified the enzyme by cobalt ion affinity chromatography. Like BCO1, purified recombinant chicken BCO2 catalyzes the oxidative cleavage of the provitamin A carotenoids β-carotene, α-carotene, and β-cryptoxanthin. Its catalytic activity with β-carotene as substrate is at least 10-fold lower than that of BCO1. In further contrast to BCO1, purified recombinant chicken BCO2 also catalyzes the oxidative cleavage of 9-cis-β-carotene and the non-provitamin A carotenoids zeaxanthin and lutein, and is inactive with all-trans-lycopene and β-apocarotenoids. Apo-10'-carotenoids were detected as enzymatic products by HPLC, and the identities were confirmed by LC-MS. Small amounts of 3-hydroxy-β-apo-8'-carotenal were also consistently detected in BCO2-β-cryptoxanthin reaction mixtures. With the exception of this activity with β-cryptoxanthin, BCO2 cleaves specifically at the 9'-10' bond to produce apo-10'-carotenoids. BCO2 has been shown to function in preventing the excessive accumulation of carotenoids, and its broad substrate specificity is consistent with this., (© 2016 by The American Society for Biochemistry and Molecular Biology, Inc.)
- Published
- 2016
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17. A Potent, Selective, and Cell-Active Inhibitor of Human Type I Protein Arginine Methyltransferases.
- Author
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Eram MS, Shen Y, Szewczyk M, Wu H, Senisterra G, Li F, Butler KV, Kaniskan HÜ, Speed BA, Dela Seña C, Dong A, Zeng H, Schapira M, Brown PJ, Arrowsmith CH, Barsyte-Lovejoy D, Liu J, Vedadi M, and Jin J
- Subjects
- Antineoplastic Agents chemistry, Cell Line, Tumor, Cell Proliferation drug effects, Ethanolamines chemistry, Ethylenediamines chemistry, Humans, Models, Molecular, Molecular Structure, Protein-Arginine N-Methyltransferases genetics, Protein-Arginine N-Methyltransferases metabolism, Pyrroles chemistry, Antineoplastic Agents pharmacology, Ethanolamines pharmacology, Ethylenediamines pharmacology, Protein-Arginine N-Methyltransferases antagonists & inhibitors, Pyrroles pharmacology
- Abstract
Protein arginine methyltransferases (PRMTs) play a crucial role in a variety of biological processes. Overexpression of PRMTs has been implicated in various human diseases including cancer. Consequently, selective small-molecule inhibitors of PRMTs have been pursued by both academia and the pharmaceutical industry as chemical tools for testing biological and therapeutic hypotheses. PRMTs are divided into three categories: type I PRMTs which catalyze mono- and asymmetric dimethylation of arginine residues, type II PRMTs which catalyze mono- and symmetric dimethylation of arginine residues, and type III PRMT which catalyzes only monomethylation of arginine residues. Here, we report the discovery of a potent, selective, and cell-active inhibitor of human type I PRMTs, MS023, and characterization of this inhibitor in a battery of biochemical, biophysical, and cellular assays. MS023 displayed high potency for type I PRMTs including PRMT1, -3, -4, -6, and -8 but was completely inactive against type II and type III PRMTs, protein lysine methyltransferases and DNA methyltransferases. A crystal structure of PRMT6 in complex with MS023 revealed that MS023 binds the substrate binding site. MS023 potently decreased cellular levels of histone arginine asymmetric dimethylation. It also reduced global levels of arginine asymmetric dimethylation and concurrently increased levels of arginine monomethylation and symmetric dimethylation in cells. We also developed MS094, a close analog of MS023, which was inactive in biochemical and cellular assays, as a negative control for chemical biology studies. MS023 and MS094 are useful chemical tools for investigating the role of type I PRMTs in health and disease.
- Published
- 2016
- Full Text
- View/download PDF
18. The human enzyme that converts dietary provitamin A carotenoids to vitamin A is a dioxygenase.
- Author
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dela Seña C, Riedl KM, Narayanasamy S, Curley RW Jr, Schwartz SJ, and Harrison EH
- Subjects
- Dioxygenases genetics, Dioxygenases metabolism, Humans, Oxygen metabolism, Recombinant Proteins chemistry, Recombinant Proteins genetics, Recombinant Proteins metabolism, Retinaldehyde genetics, Retinaldehyde metabolism, Vitamin A chemistry, Vitamin A genetics, Dioxygenases chemistry, Oxygen chemistry, Retinaldehyde chemistry, Vitamin A biosynthesis
- Abstract
β-Carotene 15-15'-oxygenase (BCO1) catalyzes the oxidative cleavage of dietary provitamin A carotenoids to retinal (vitamin A aldehyde). Aldehydes readily exchange their carbonyl oxygen with water, making oxygen labeling experiments challenging. BCO1 has been thought to be a monooxygenase, incorporating oxygen from O2 and H2O into its cleavage products. This was based on a study that used conditions that favored oxygen exchange with water. We incubated purified recombinant human BCO1 and β-carotene in either (16)O2-H2(18)O or (18)O2-H2(16)O medium for 15 min at 37 °C, and the relative amounts of (18)O-retinal and (16)O-retinal were measured by liquid chromatography-tandem mass spectrometry. At least 79% of the retinal produced by the reaction has the same oxygen isotope as the O2 gas used. Together with the data from (18)O-retinal-H2(16)O and (16)O-retinal-H2(18)O incubations to account for nonenzymatic oxygen exchange, our results show that BCO1 incorporates only oxygen from O2 into retinal. Thus, BCO1 is a dioxygenase.
- Published
- 2014
- Full Text
- View/download PDF
19. Substrate specificity of purified recombinant human β-carotene 15,15'-oxygenase (BCO1).
- Author
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dela Seña C, Narayanasamy S, Riedl KM, Curley RW Jr, Schwartz SJ, and Harrison EH
- Subjects
- Carotenoids metabolism, Catalysis, Humans, Oxidation-Reduction, Recombinant Proteins chemistry, Recombinant Proteins genetics, Recombinant Proteins isolation & purification, Recombinant Proteins metabolism, Substrate Specificity, beta-Carotene 15,15'-Monooxygenase genetics, beta-Carotene 15,15'-Monooxygenase isolation & purification, beta-Carotene 15,15'-Monooxygenase metabolism, Carotenoids chemistry, beta-Carotene 15,15'-Monooxygenase chemistry
- Abstract
Humans cannot synthesize vitamin A and thus must obtain it from their diet. β-Carotene 15,15'-oxygenase (BCO1) catalyzes the oxidative cleavage of provitamin A carotenoids at the central 15-15' double bond to yield retinal (vitamin A). In this work, we quantitatively describe the substrate specificity of purified recombinant human BCO1 in terms of catalytic efficiency values (kcat/Km). The full-length open reading frame of human BCO1 was cloned into the pET-28b expression vector with a C-terminal polyhistidine tag, and the protein was expressed in the Escherichia coli strain BL21-Gold(DE3). The enzyme was purified using cobalt ion affinity chromatography. The purified enzyme preparation catalyzed the oxidative cleavage of β-carotene with a Vmax = 197.2 nmol retinal/mg BCO1 × h, Km = 17.2 μM and catalytic efficiency kcat/Km = 6098 M(-1) min(-1). The enzyme also catalyzed the oxidative cleavage of α-carotene, β-cryptoxanthin, and β-apo-8'-carotenal to yield retinal. The catalytic efficiency values of these substrates are lower than that of β-carotene. Surprisingly, BCO1 catalyzed the oxidative cleavage of lycopene to yield acycloretinal with a catalytic efficiency similar to that of β-carotene. The shorter β-apocarotenals (β-apo-10'-carotenal, β-apo-12'-carotenal, β-apo-14'-carotenal) do not show Michaelis-Menten behavior under the conditions tested. We did not detect any activity with lutein, zeaxanthin, and 9-cis-β-carotene. Our results show that BCO1 favors full-length provitamin A carotenoids as substrates, with the notable exception of lycopene. Lycopene has previously been reported to be unreactive with BCO1, and our findings warrant a fresh look at acycloretinal and its alcohol and acid forms as metabolites of lycopene in future studies.
- Published
- 2013
- Full Text
- View/download PDF
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