40 results on '"Faust B"'
Search Results
2. Numerical modelling of the compaction behaviour of crushed rock salt
- Author
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Heemann, U., primary, Heusermann, S., additional, Sarfeld, W., additional, and Faust, B., additional
- Published
- 2020
- Full Text
- View/download PDF
3. Calibration of angle standards
- Author
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Gastaldi, B, primary, Vieira, L, additional, Yshikawa, F Huerta, additional, Faust, B, additional, and Eves, B, additional
- Published
- 2023
- Full Text
- View/download PDF
4. Crack-initiation and propagation in rock salt under hydromechanical interaction
- Author
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Heemann, U., primary, Sarfeld, W., additional, Hillmann, C., additional, and Faust, B., additional
- Published
- 2017
- Full Text
- View/download PDF
5. Human Thyrotropin receptor bound by CS-17 Inverse Agonist Fab/Org 274179-0 Antagonist
- Author
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Faust, B., primary, Cheng, Y., additional, and Manglik, A., additional
- Published
- 2022
- Full Text
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6. Human thyrotropin analog TR1402 bound to human Thyrotropin receptor in complex with miniGs399 (composite structure)
- Author
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Faust, B., primary, Cheng, Y., additional, and Manglik, A., additional
- Published
- 2022
- Full Text
- View/download PDF
7. M22 Agonist Autoantibody bound to Human Thyrotropin receptor in complex with miniGs399 (composite structure)
- Author
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Faust, B., primary, Cheng, Y., additional, and Manglik, A., additional
- Published
- 2022
- Full Text
- View/download PDF
8. Native human TSH bound to human Thyrotropin receptor in complex with miniGs399 (composite structure)
- Author
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Faust, B., primary, Cheng, Y., additional, and Manglik, A., additional
- Published
- 2022
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- View/download PDF
9. Substance P bound to active human neurokinin 1 receptor in complex with miniGs399
- Author
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Harris, J.A., primary, Faust, B., additional, Gondin, A.B., additional, Daemgen, M.A., additional, Suomivuori, C.M., additional, Veldhuis, N.A., additional, Cheng, Y., additional, Dror, R.O., additional, Thal, D., additional, and Manglik, A., additional
- Published
- 2021
- Full Text
- View/download PDF
10. SP6-11 biased agonist bound to active human neurokinin 1 receptor in complex with miniGs/q70
- Author
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Harris, J.A., primary, Faust, B., additional, Gondin, A.B., additional, Daemgen, M.A., additional, Suomivuori, C.M., additional, Veldhuis, N.A., additional, Cheng, Y., additional, Dror, R.O., additional, Thal, D., additional, and Manglik, A., additional
- Published
- 2021
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- View/download PDF
11. Functional and structural studies of the human ABC transporter ABCB8
- Author
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Faust, B, Carpenter, E, and Shintre, C
- Abstract
ATP-binding cassette (ABC) transporters are one of the largest families of membrane proteins and are ATP-driven transport systems, which are found universally in all pro-and eukaryotes. Human ABC transporters have key roles in many physiological processes in the plasma membrane and within membranes of cell organelles, such as peptide transport, lipid flipping and multi-drug resistance. ABCB8, a sub-family B ABC transporter of the inner mitochondrial membrane, was found to be crucial for normal cardiac function, regulation of mitochondrial iron levels and cytosolic iron-sulphur cluster enzyme function and oxidative stress. Impaired function of ABCB8 resulted in mitochondrial iron accumulation, oxidative stress and cell death. In this thesis I developed a protocol for the purification of human ABCB8 pro¬viding yields and stable protein sufficient for structural studies using X-ray crystallography. Strategic optimisation of the purification protocol and crystallisation methods, resulted in crystals that diffracted to a resolution of 3.8 Å and allowed me to solve the first structure of human ABCB8. The ABCB8 structure adopted an outward-facing conformation that resembles a post-ATP-hydrolysis state in the alternating access mechanism, that was not yet observed in type I ABC exporters. In addition to the structural studies on ABCB8, I set out to explore the effects of ATP analogues, ATP transition state analogues and ADP on purified ABCB8 with biochemical and biophysical methods. Furthermore, I find that ATPase activity of ABCB8 in proteoliposomes is dependent on cardiolipin (CDL). The first structure of ABCB8 in an outward-facing conformation gives new insights into the alternating-access mechanism of subfamily B ABC transporters. Moreover, the purification and reconstitution protocol provided in this thesis is the foundation for further studies aiming to understand ABCB8’s function in more detail.
- Published
- 2018
12. Crystal structure of active Smoothened bound to SAG21k, cholesterol, and NbSmo8
- Author
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Deshpande, I.S., primary, Liang, J., additional, Hedeen, D., additional, Roberts, K.J., additional, Zhang, Y., additional, Ha, B., additional, Latorraca, N.R., additional, Faust, B., additional, Dror, R.O., additional, Beachy, P.A., additional, Myers, B.R., additional, and Manglik, A., additional
- Published
- 2019
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13. Human m7GpppN-mRNA Hydrolase (DCP2, NUDT20) Catalytic Domain
- Author
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Mathea, S., primary, Salah, E., additional, Velupillai, S., additional, Tallant, C., additional, Pike, A.C.W., additional, Bushell, S.R., additional, Faust, B., additional, Wang, D., additional, Burgess-Brown, N., additional, von Delft, F., additional, Arrowsmith, C.H., additional, Edwards, A.M., additional, Bountra, C., additional, Knapp, S., additional, and Huber, K., additional
- Published
- 2017
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14. Multidimensional Poverty Indicators in light of ASEAN Integration
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Jayvee Faust B. Anga, Kristine June D. Uy, Jonah Marie S. Enerlas, and Marjurie Lourince E. Zanoria
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Economic integration ,H1-99 ,Index (economics) ,Poverty ,HF5001-6182 ,poverty ,Corporate governance ,media_common.quotation_subject ,General Engineering ,Disease cluster ,asean ,Multidimensional Poverty Index ,asean integration ,Social sciences (General) ,Investment decisions ,Geography ,Development economics ,T1-995 ,Business ,Welfare ,Technology (General) ,media_common - Abstract
This study aimed to explore poverty among ASEAN member nations by introducing a multidimensional poverty index that can be used to determine the relative success of a country in deterring several deprivations that are identified and measured. It utilized multiple factors that can be useful for policy and investment decisions and are intended to complement analyses using financial poverty indicators. Using exploratory data analysis, several indicators were selected to represent the different dimensions of poverty. Said indicators were then subjected to factor analysis which yielded four multidimensional indices namely: General Welfare Index, Governance and Emotional Landscape Index, and Social Climate Index. Singapore, Brunei, and Malaysia ranked 1st, 2nd, and 3rd in all three indices, respectively. In addition, multivariate cluster analysis was also performed. It revealed that Singapore exhibited exceptional performance in all indices, thereby earning it a cluster all to its own. Similarities were observed between Brunei and Malaysia while Thailand, Vietnam, Philippines, and Indonesia were clustered together. Cambodia, Lao PDR, and Myanmar belonged to another cluster, indicating similarities among them. The study concluded that a wide gap exists among ASEAN countries in terms of the various deprivations measured. It also identified several problem areas and strengths of the ASEAN member states. This information can be helpful in making sound judgments especially in the delicate circumstances that surround economic integration.
- Published
- 2015
15. Crystal structure of the BTB domain of human KCTD17
- Author
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Pinkas, D.M., primary, Sorrell, F., additional, Sanvitale, C.E., additional, Goubin, S., additional, Williams, E., additional, Newman, J.A., additional, Pearce, N.M., additional, Neshich, I., additional, Pike, A.C.W., additional, MacKenzie, A., additional, Quigley, A., additional, Faust, B., additional, Carpenter, E.P., additional, Tallant, C., additional, Kopec, J., additional, Chalk, R., additional, Krojer, T., additional, Burgess-Brown, N.A., additional, von Delft, F., additional, Arrowsmith, C.H., additional, Edwards, A.M., additional, Bountra, C., additional, and Bullock, A., additional
- Published
- 2015
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16. Structural characterization of ligand binding and pH-specific enzymatic activity of mouse Acidic Mammalian Chitinase.
- Author
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Díaz RE, Ecker AK, Correy GJ, Asthana P, Young ID, Faust B, Thompson MC, Seiple IB, Van Dyken S, Locksley RM, and Fraser JS
- Subjects
- Animals, Hydrogen-Ion Concentration, Mice, Protein Conformation, Crystallography, X-Ray, Protein Binding, Ligands, Kinetics, Acetylglucosamine metabolism, Acetylglucosamine chemistry, Models, Molecular, Chitinases metabolism, Chitinases chemistry, Molecular Dynamics Simulation, Chitin metabolism, Chitin chemistry
- Abstract
Chitin is an abundant biopolymer and pathogen-associated molecular pattern that stimulates a host innate immune response. Mammals express chitin-binding and chitin-degrading proteins to remove chitin from the body. One of these proteins, Acidic Mammalian Chitinase (AMCase), is an enzyme known for its ability to function under acidic conditions in the stomach but is also active in tissues with more neutral pHs, such as the lung. Here, we used a combination of biochemical, structural, and computational modeling approaches to examine how the mouse homolog (mAMCase) can act in both acidic and neutral environments. We measured kinetic properties of mAMCase activity across a broad pH range, quantifying its unusual dual activity optima at pH 2 and 7. We also solved high-resolution crystal structures of mAMCase in complex with oligomeric GlcNAcn, the building block of chitin, where we identified extensive conformational ligand heterogeneity. Leveraging these data, we conducted molecular dynamics simulations that suggest how a key catalytic residue could be protonated via distinct mechanisms in each of the two environmental pH ranges. These results integrate structural, biochemical, and computational approaches to deliver a more complete understanding of the catalytic mechanism governing mAMCase activity at different pH. Engineering proteins with tunable pH optima may provide new opportunities to develop improved enzyme variants, including AMCase, for therapeutic purposes in chitin degradation., Competing Interests: RD, AE, GC, PA, IY, BF, MT, IS No competing interests declared, SV, RL S.J.V.D. and R.M.L. are listed as inventors on a patent United States application: 17/505,561 for the use of chitinases to treat fibrotic lung disease. S.J.V.D., R.M.L., and J.S.F. are listed as inventors on a patent for mutant chitinases with enhanced expression and activity, JF S.J.V.D. and R.M.L. are listed as inventors on a patent for the use of chitinases to treat fibrotic lung disease. United States application: 17/505,561 S.J.V.D., R.M.L., and J.S.F. are listed as inventors on a patent for mutant chitinases with enhanced expression and activity, (© 2023, Díaz et al.)
- Published
- 2024
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17. Engineered odorant receptors illuminate structural principles of odor discrimination.
- Author
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de March CA, Ma N, Billesbølle CB, Tewari J, Del Torrent CL, van der Velden WJC, Ojiro I, Takayama I, Faust B, Li L, Vaidehi N, Manglik A, and Matsunami H
- Abstract
A central challenge in olfaction is understanding how the olfactory system detects and distinguishes odorants with diverse physicochemical properties and molecular configurations. Vertebrate animals perceive odors via G protein-coupled odorant receptors (ORs). In humans, ~400 ORs enable the sense of smell. The OR family is composed of two major classes: Class I ORs are tuned to carboxylic acids while Class II ORs, representing the vast majority of the human repertoire, respond to a wide variety of odorants. How ORs recognize chemically diverse odorants remains poorly understood. A fundamental bottleneck is the inability to visualize odorant binding to ORs. Here, we uncover fundamental molecular properties of odorant-OR interactions by employing engineered ORs crafted using a consensus protein design strategy. Because such consensus ORs (consORs) are derived from the 17 major subfamilies of human ORs, they provide a template for modeling individual native ORs with high sequence and structural homology. The biochemical tractability of consORs enabled four cryoEM structures of distinct consORs with unique ligand recognition properties. The structure of a Class I consOR, consOR51, showed high structural similarity to the native human receptor OR51E2 and yielded a homology model of a related member of the human OR51 family with high predictive power. Structures of three Class II consORs revealed distinct modes of odorant-binding and activation mechanisms between Class I and Class II ORs. Thus, the structures of consORs lay the groundwork for understanding molecular recognition of odorants by the OR superfamily., Competing Interests: H.M. has received royalties from Chemcom, research grants from Givaudan, and consultant fees from Kao. A.M. is a founder of Epiodyne and Stipple Bio, consults for Abalone, and serves on the scientific advisory board of Septerna.
- Published
- 2023
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18. Structural basis of odorant recognition by a human odorant receptor.
- Author
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Billesbølle CB, de March CA, van der Velden WJC, Ma N, Tewari J, Del Torrent CL, Li L, Faust B, Vaidehi N, Matsunami H, and Manglik A
- Subjects
- Humans, Smell physiology, Molecular Dynamics Simulation, Mutation, Binding Sites genetics, Substrate Specificity genetics, Cryoelectron Microscopy, Odorants analysis, Propionates chemistry, Propionates metabolism, Receptors, Odorant chemistry, Receptors, Odorant genetics, Receptors, Odorant metabolism, Receptors, Odorant ultrastructure
- Abstract
Our sense of smell enables us to navigate a vast space of chemically diverse odour molecules. This task is accomplished by the combinatorial activation of approximately 400 odorant G protein-coupled receptors encoded in the human genome
1-3 . How odorants are recognized by odorant receptors remains unclear. Here we provide mechanistic insight into how an odorant binds to a human odorant receptor. Using cryo-electron microscopy, we determined the structure of the active human odorant receptor OR51E2 bound to the fatty acid propionate. Propionate is bound within an occluded pocket in OR51E2 and makes specific contacts critical to receptor activation. Mutation of the odorant-binding pocket in OR51E2 alters the recognition spectrum for fatty acids of varying chain length, suggesting that odorant selectivity is controlled by tight packing interactions between an odorant and an odorant receptor. Molecular dynamics simulations demonstrate that propionate-induced conformational changes in extracellular loop 3 activate OR51E2. Together, our studies provide a high-resolution view of chemical recognition of an odorant by a vertebrate odorant receptor, providing insight into how this large family of G protein-coupled receptors enables our olfactory sense., (© 2023. The Author(s), under exclusive licence to Springer Nature Limited.)- Published
- 2023
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19. Autoantibody mimicry of hormone action at the thyrotropin receptor.
- Author
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Faust B, Billesbølle CB, Suomivuori CM, Singh I, Zhang K, Hoppe N, Pinto AFM, Diedrich JK, Muftuoglu Y, Szkudlinski MW, Saghatelian A, Dror RO, Cheng Y, and Manglik A
- Subjects
- Cell Membrane metabolism, Graves Disease immunology, Graves Disease metabolism, Humans, Phospholipids metabolism, Protein Domains, Receptors, G-Protein-Coupled agonists, Receptors, G-Protein-Coupled chemistry, Receptors, G-Protein-Coupled ultrastructure, Rotation, Cryoelectron Microscopy, Immunoglobulins, Thyroid-Stimulating chemistry, Immunoglobulins, Thyroid-Stimulating immunology, Immunoglobulins, Thyroid-Stimulating pharmacology, Immunoglobulins, Thyroid-Stimulating ultrastructure, Receptors, Thyrotropin agonists, Receptors, Thyrotropin chemistry, Receptors, Thyrotropin immunology, Receptors, Thyrotropin ultrastructure, Thyrotropin chemistry, Thyrotropin metabolism, Thyrotropin pharmacology
- Abstract
Thyroid hormones are vital in metabolism, growth and development
1 . Thyroid hormone synthesis is controlled by thyrotropin (TSH), which acts at the thyrotropin receptor (TSHR)2 . In patients with Graves' disease, autoantibodies that activate the TSHR pathologically increase thyroid hormone activity3 . How autoantibodies mimic thyrotropin function remains unclear. Here we determined cryo-electron microscopy structures of active and inactive TSHR. In inactive TSHR, the extracellular domain lies close to the membrane bilayer. Thyrotropin selects an upright orientation of the extracellular domain owing to steric clashes between a conserved hormone glycan and the membrane bilayer. An activating autoantibody from a patient with Graves' disease selects a similar upright orientation of the extracellular domain. Reorientation of the extracellular domain transduces a conformational change in the seven-transmembrane-segment domain via a conserved hinge domain, a tethered peptide agonist and a phospholipid that binds within the seven-transmembrane-segment domain. Rotation of the TSHR extracellular domain relative to the membrane bilayer is sufficient for receptor activation, revealing a shared mechanism for other glycoprotein hormone receptors that may also extend to other G-protein-coupled receptors with large extracellular domains., (© 2022. The Author(s), under exclusive licence to Springer Nature Limited.)- Published
- 2022
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20. The Lectin LecB Induces Patches with Basolateral Characteristics at the Apical Membrane to Promote Pseudomonas aeruginosa Host Cell Invasion.
- Author
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Thuenauer R, Kühn K, Guo Y, Kotsis F, Xu M, Trefzer A, Altmann S, Wehrum S, Heshmatpour N, Faust B, Landi A, Diedrich B, Dengjel J, Kuehn EW, Imberty A, and Römer W
- Subjects
- Actins metabolism, Caveolin 1 metabolism, Cell Membrane metabolism, Humans, Phosphatidylinositol 3-Kinases metabolism, Lectins metabolism, Pseudomonas aeruginosa metabolism
- Abstract
The opportunistic bacterium Pseudomonas aeruginosa can infect mucosal tissues of the human body. To persist at the mucosal barrier, this highly adaptable pathogen has evolved many strategies, including invasion of host cells. Here, we show that the P. aeruginosa lectin LecB binds and cross-links fucosylated receptors at the apical plasma membrane of epithelial cells. This triggers a signaling cascade via Src kinases and phosphoinositide 3-kinase (PI3K), leading to the formation of patches enriched with the basolateral marker phosphatidylinositol (3,4,5)-trisphosphate (PIP
3 ) at the apical plasma membrane. This identifies LecB as a causative bacterial factor for activating this well-known host cell response that is elicited upon apical binding of P. aeruginosa. Downstream from PI3K, Rac1 is activated to cause actin rearrangement and the outgrowth of protrusions at the apical plasma membrane. LecB-triggered PI3K activation also results in aberrant recruitment of caveolin-1 to the apical domain. In addition, we reveal a positive feedback loop between PI3K activation and apical caveolin-1 recruitment, which provides a mechanistic explanation for the previously observed implication of caveolin-1 in P. aeruginosa host cell invasion. Interestingly, LecB treatment also reversibly removes primary cilia. To directly prove the role of LecB for bacterial uptake, we coated bacterium-sized beads with LecB, which drastically enhanced their endocytosis. Furthermore, LecB deletion and LecB inhibition with l-fucose diminished the invasion efficiency of P. aeruginosa bacteria. Taken together, the results of our study identify LecB as a missing link that can explain how PI3K signaling and caveolin-1 recruitment are triggered to facilitate invasion of epithelial cells from the apical side by P. aeruginosa. IMPORTANCE An intriguing feature of the bacterium P. aeruginosa is its ability to colonize highly diverse niches. P. aeruginosa can, besides forming biofilms, also enter and proliferate within epithelial host cells. Moreover, research during recent years has shown that P. aeruginosa possesses many different mechanisms to invade host cells. In this study, we identify LecB as a novel invasion factor. In particular, we show that LecB activates PI3K signaling, which is connected via a positive feedback loop to apical caveolin-1 recruitment and leads to actin rearrangement at the apical plasma membrane. This provides a unifying explanation for the previously reported implication of PI3K and caveolin-1 in host cell invasion by P. aeruginosa. In addition, our study adds a further function to the remarkable repertoire of the lectin LecB, which is all brought about by the capability of LecB to recognize fucosylated glycans on many different niche-specific host cell receptors.- Published
- 2022
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21. Pharmacology update: pamidronate for hypertrophic pulmonary osteoarthropathy in palliative care.
- Author
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Faust B, Parkinson A, and Baumrucker SJ
- Abstract
Hypertrophic pulmonary osteoarthropathy (HPOA) is a rare syndrome that causes clubbed fingers, periostitis, and synovial effusions. It can adversely impact a patient's quality of life. It occurs secondary to pulmonary disease - most commonly pulmonary malignancy. The most effective treatment for HPOA is to treat the underlying disease, usually through surgical resection, chemotherapy, or radiation. However, symptomatic treatments rather than definitive treatments (surgical, chemotherapy, or radiation) are more appropriate for the palliative care patient. Pamidronate is a promising medication for the treatment of HPOA for its safety and rapid onset of action. Further research is indicated to determine whether pamidronate is consistently effective., Competing Interests: The authors declared no potential conflicts of interest with respect to the research, authorship, and/or publication of this article., (© The Author(s), 2022.)
- Published
- 2022
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22. Guidance in selecting analytical techniques for identification and quantification of non-intentionally added substances (NIAS) in food contact materials (FCMS).
- Author
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Nerín C, Bourdoux S, Faust B, Gude T, Lesueur C, Simat T, Stoermer A, Van Hoek E, and Oldring P
- Subjects
- Allergens analysis, Food, Food Contamination analysis, Food Packaging
- Abstract
There are numerous approaches and methodologies for assessing the identity and quantities of non-intentionally added substances (NIAS) in food contact materials (FCMs). They can give different results and it can be difficult to make meaningful comparisons. The initial approach was to attempt to prepare a prescriptive methodology but as this proved impossible; this paper develops guidelines that need to be taken into consideration when assessing NIAS. Different approaches to analysing NIAS in FCMs are reviewed and compared. The approaches for preparing the sample for analysis, recommended procedures for screening, identification, and quantification of NIAS as well as the reporting requirements are outlined. Different analytical equipment and procedures are compared. Limitations of today's capabilities are raised along with some research needs.
- Published
- 2022
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23. Divergence Between Informant and Self-Ratings of Activities of Daily Living Impairments in Parkinson's Disease.
- Author
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Becker S, Solbrig S, Michaelis K, Faust B, Brockmann K, and Liepelt-Scarfone I
- Abstract
Objective: To examine the agreement between self- and informant-reported activities of daily living (ADL) deficits in Parkinson's Disease (PD) patients, and to examine factors influencing ADL ratings., Background: In PD, the loss of functional independence is an important outcome of disease progression. The valid assessment of ADL function in PD is essential, but it is unclear to what extent informants' and patients' perceptions of their daily functions concur, and how other factors may influence both ratings., Methods: Data of 150 PD patients who underwent cognitive and motor testing, as well as their informants were analyzed. The 10-item Functional Activities Questionnaire (FAQ), completed separately by patients (FAQ-S) and their informants (FAQ-I), assessed ADL function. Weighted κ statistics summarized level of agreement, and a discrepancy score (FAQ-I - FAQ-S) quantified agreement. Correlation analyses between FAQ total scores, patient and informant characteristics, and cognitive scores were conducted, with post hoc regressions to determine the associations between both FAQ scores and cognition, independent of patient characteristics., Results: The sample included 87 patients with normal cognition, 50 with mild cognitive impairment, and 13 with dementia. Overall, there was fair to moderate agreement between patients and informants on individual FAQ items (0.27 ≤ κ ≤ 0.61, p < 0.004), with greater discrepancies with increasing cognitive impairment. Patients' age, motor severity, non-motor burden, and depression also affected both ratings (0.27 ≤ r ≤ 0.50, p < 0.001), with motor severity showing the greatest influence on both ratings. Both the FAQ-I and FAQ-S were correlated with almost all cognitive domains. Post hoc regression analyses controlling for patient characteristics showed that the attention domain was a significant predictor of both the FAQ-S and FAQ-I scores, and memory was also a significant predictor of the FAQ-I score. Only 29.3% of patients agreed perfectly with informants on the FAQ total score, with informants most commonly rating ADL impairments as more severe than patients., Conclusions: Patient and informant ratings of ADL function using FAQ items showed moderate agreement, with only few items reaching substantial agreement. Ratings of both were associated with patient cognitive status, but also other characteristics. In addition to patient and informant reports, objective measures are needed to accurately classify ADL deficits in PD., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2022 Becker, Solbrig, Michaelis, Faust, Brockmann and Liepelt-Scarfone.)
- Published
- 2022
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24. Selective G protein signaling driven by substance P-neurokinin receptor dynamics.
- Author
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Harris JA, Faust B, Gondin AB, Dämgen MA, Suomivuori CM, Veldhuis NA, Cheng Y, Dror RO, Thal DM, and Manglik A
- Subjects
- Animals, Inflammation metabolism, GTP-Binding Proteins metabolism, Receptors, Neurokinin-1 metabolism, Signal Transduction
- Abstract
The neuropeptide substance P (SP) is important in pain and inflammation. SP activates the neurokinin-1 receptor (NK1R) to signal via G
q and Gs proteins. Neurokinin A also activates NK1R, but leads to selective Gq signaling. How two stimuli yield distinct G protein signaling at the same G protein-coupled receptor remains unclear. We determined cryogenic-electron microscopy structures of active NK1R bound to SP or the Gq -biased peptide SP6-11. Peptide interactions deep within NK1R are critical for receptor activation. Conversely, interactions between SP and NK1R extracellular loops are required for potent Gs signaling but not Gq signaling. Molecular dynamics simulations showed that these superficial contacts restrict SP flexibility. SP6-11, which lacks these interactions, is dynamic while bound to NK1R. Structural dynamics of NK1R agonists therefore depend on interactions with the receptor extracellular loops and regulate G protein signaling selectivity. Similar interactions between other neuropeptides and their cognate receptors may tune intracellular signaling., (© 2021. The Author(s), under exclusive licence to Springer Nature America, Inc.)- Published
- 2022
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25. Structural insight into SARS-CoV-2 neutralizing antibodies and modulation of syncytia.
- Author
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Asarnow D, Wang B, Lee WH, Hu Y, Huang CW, Faust B, Ng PML, Ngoh EZX, Bohn M, Bulkley D, Pizzorno A, Ary B, Tan HC, Lee CY, Minhat RA, Terrier O, Soh MK, Teo FJ, Yeap YYC, Seah SGK, Chan CEZ, Connelly E, Young NJ, Maurer-Stroh S, Renia L, Hanson BJ, Rosa-Calatrava M, Manglik A, Cheng Y, Craik CS, and Wang CI
- Subjects
- Angiotensin-Converting Enzyme 2 chemistry, Angiotensin-Converting Enzyme 2 genetics, Angiotensin-Converting Enzyme 2 immunology, Angiotensin-Converting Enzyme 2 metabolism, Animals, Antibodies, Neutralizing immunology, Antibodies, Neutralizing metabolism, Antigen-Antibody Complex chemistry, Antigen-Antibody Complex metabolism, Binding Sites, CHO Cells, COVID-19 pathology, COVID-19 virology, Cricetinae, Cricetulus, Cryoelectron Microscopy, Giant Cells cytology, Humans, Membrane Fusion, Peptide Library, Protein Binding, Protein Domains, Protein Structure, Quaternary, SARS-CoV-2 isolation & purification, SARS-CoV-2 metabolism, Spike Glycoprotein, Coronavirus immunology, Spike Glycoprotein, Coronavirus metabolism, Antibodies, Neutralizing chemistry, Giant Cells metabolism, Spike Glycoprotein, Coronavirus chemistry
- Abstract
Infection with severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) is initiated by binding of the viral Spike protein to host receptor angiotensin-converting enzyme 2 (ACE2), followed by fusion of viral and host membranes. Although antibodies that block this interaction are in emergency use as early coronavirus disease 2019 (COVID-19) therapies, the precise determinants of neutralization potency remain unknown. We discovered a series of antibodies that potently block ACE2 binding but exhibit divergent neutralization efficacy against the live virus. Strikingly, these neutralizing antibodies can inhibit or enhance Spike-mediated membrane fusion and formation of syncytia, which are associated with chronic tissue damage in individuals with COVID-19. As revealed by cryoelectron microscopy, multiple structures of Spike-antibody complexes have distinct binding modes that not only block ACE2 binding but also alter the Spike protein conformational cycle triggered by ACE2 binding. We show that stabilization of different Spike conformations leads to modulation of Spike-mediated membrane fusion with profound implications for COVID-19 pathology and immunity., Competing Interests: Declaration of interests B.W., W.-H.L., C.-W.H., Y.H., P.M.L.N., E.Z.X.N., H.C.T., C.Y.L., R.A.M., M.K.S., F.J.T., Y.Y.C.Y., and C.-I.W. are listed as inventors of a filed patent for all 27 monoclonal antibodies mentioned in this manuscript., (Copyright © 2021 Elsevier Inc. All rights reserved.)
- Published
- 2021
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26. Pre-season in soccer: a paradox between a high volume of technical/tactical training and improvement in the neuromuscular performance of elite women soccer players.
- Author
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Kobal R, Carvalho L, Abad CC, Faust B, Rossetti M, Saito T, Klosterhoff RR, De Souza EO, and Barroso R
- Subjects
- Adipose Tissue, Female, Humans, Muscle Strength, Seasons, Athletic Performance, Resistance Training methods, Soccer
- Abstract
Background: there is a paradox between the development of strength-power abilities and the high volume of technical/tactical training in elite soccer players during the pre-season. This concurrent effect between aerobic and neuromuscular training regimes induce impairment in power performance., Methods: this study aimed to investigate the effect of an equalized program of strength-power training (4-5 sessions/week) and soccer training (4-6 sessions/week) in power and aerobic performance during 8-weeks of pre-season in elite women soccer players. Vertical jumps [squat jump (SJ); countermovement jump (CMJ)] and Yo-Yo intermittent recovery level 1 test (YOYO-R1) were assessed pre- and post pre-season. A paired sample t-test was used to compare differences between pre and post pre-season (Δ%). The level of significance was established at p ≤ 0.05., Results: the women soccer players improved the SJ (p<0.001; Δ%=12), CMJ (p<0.001; Δ%=8.5), and YOYO-R1 (p<0.001; Δ% =28.5). There was a body recomposition observed, lower body fat (p = 0.004; Δ%=15), higher fat free mass (p = 0.001; Δ%=5)., Conclusions: our results demonstrated that it is possible to develop aerobic and power abilities of elite women soccer players during pre-season using an equalized ratio of soccer training and strength-power training schedules.
- Published
- 2021
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27. ABCB10 exports mitochondrial biliverdin, driving metabolic maladaptation in obesity.
- Author
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Shum M, Shintre CA, Althoff T, Gutierrez V, Segawa M, Saxberg AD, Martinez M, Adamson R, Young MR, Faust B, Gharakhanian R, Su S, Chella Krishnan K, Mahdaviani K, Veliova M, Wolf DM, Ngo J, Nocito L, Stiles L, Abramson J, Lusis AJ, Hevener AL, Zoghbi ME, Carpenter EP, and Liesa M
- Subjects
- Animals, Antioxidants, Bilirubin, Liver, Mice, Obesity, Biliverdine, Mitochondria
- Abstract
Although the role of hydrophilic antioxidants in the development of hepatic insulin resistance and nonalcoholic fatty liver disease has been well studied, the role of lipophilic antioxidants remains poorly characterized. A known lipophilic hydrogen peroxide scavenger is bilirubin, which can be oxidized to biliverdin and then reduced back to bilirubin by cytosolic biliverdin reductase. Oxidation of bilirubin to biliverdin inside mitochondria must be followed by the export of biliverdin to the cytosol, where biliverdin is reduced back to bilirubin. Thus, the putative mitochondrial exporter of biliverdin is expected to be a major determinant of bilirubin regeneration and intracellular hydrogen peroxide scavenging. Here, we identified ABCB10 as a mitochondrial biliverdin exporter. ABCB10 reconstituted into liposomes transported biliverdin, and ABCB10 deletion caused accumulation of biliverdin inside mitochondria. Obesity with insulin resistance up-regulated hepatic ABCB10 expression in mice and elevated cytosolic and mitochondrial bilirubin content in an ABCB10-dependent manner. Revealing a maladaptive role of ABCB10-driven bilirubin synthesis, hepatic ABCB10 deletion protected diet-induced obese mice from steatosis and hyperglycemia, improving insulin-mediated suppression of glucose production and decreasing lipogenic SREBP-1c expression. Protection was concurrent with enhanced mitochondrial function and increased inactivation of PTP1B, a phosphatase disrupting insulin signaling and elevating SREBP-1c expression. Restoration of cellular bilirubin content in ABCB10 KO hepatocytes reversed the improvements in mitochondrial function and PTP1B inactivation, demonstrating that bilirubin was the maladaptive effector linked to ABCB10 function. Thus, we identified a fundamental transport process that amplifies intracellular bilirubin redox actions, which can exacerbate insulin resistance and steatosis in obesity., (Copyright © 2021 The Authors, some rights reserved; exclusive licensee American Association for the Advancement of Science. No claim to original U.S. Government Works.)
- Published
- 2021
- Full Text
- View/download PDF
28. Complex of human Melanotransferrin and SC57.32 Fab fragment reveals novel interdomain arrangement with ferric N-lobe and open C-lobe.
- Author
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Hayashi K, Longenecker KL, Liu YL, Faust B, Prashar A, Hampl J, Stoll V, and Vivona S
- Subjects
- Acetylglucosamine, Animals, Biological Transport, Blood-Brain Barrier metabolism, Drug Delivery Systems, Drug Design, Gene Expression, Humans, Immunoglobulin Fab Fragments genetics, Immunoglobulin Fab Fragments metabolism, Immunoglobulin Fab Fragments physiology, Iron metabolism, Macaca fascicularis, Melanoma etiology, Melanoma genetics, Membrane Glycoproteins genetics, Membrane Glycoproteins metabolism, Mice, Protein Binding, Triple Negative Breast Neoplasms genetics, Immunoglobulin Fab Fragments chemistry, Membrane Glycoproteins chemistry, Membrane Glycoproteins physiology, Protein Domains
- Abstract
Melanotransferrin (MTf) is an iron-binding member of the transferrin superfamily that can be membrane-anchored or secreted in serum. On cells, it can mediate transferrin-independent iron uptake and promote proliferation. In serum, it is a transcytotic iron transporter across the blood-brain barrier. MTf has been exploited as a drug delivery carrier to the brain and as an antibody-drug conjugate (ADC) target due to its oncogenic role in melanoma and its elevated expression in triple-negative breast cancer (TNBC). For treatment of TNBC, an MTf-targeting ADC completed a phase I clinical trial (NCT03316794). The structure of its murine, unconjugated Fab fragment (SC57.32) is revealed here in complex with MTf. The MTf N-lobe is in an active and iron-bound, closed conformation while the C-lobe is in an open conformation incompatible with iron binding. This combination of active and inactive domains displays a novel inter-domain arrangement in which the C2 subdomain angles away from the N-lobe. The C2 subdomain also contains the SC57.32 glyco-epitope, which comprises ten protein residues and two N-acetylglucosamines. Our report reveals novel features of MTf and provides a point of reference for MTf-targeting, structure-guided drug design.
- Published
- 2021
- Full Text
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29. An ultrapotent synthetic nanobody neutralizes SARS-CoV-2 by stabilizing inactive Spike.
- Author
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Schoof M, Faust B, Saunders RA, Sangwan S, Rezelj V, Hoppe N, Boone M, Billesbølle CB, Puchades C, Azumaya CM, Kratochvil HT, Zimanyi M, Deshpande I, Liang J, Dickinson S, Nguyen HC, Chio CM, Merz GE, Thompson MC, Diwanji D, Schaefer K, Anand AA, Dobzinski N, Zha BS, Simoneau CR, Leon K, White KM, Chio US, Gupta M, Jin M, Li F, Liu Y, Zhang K, Bulkley D, Sun M, Smith AM, Rizo AN, Moss F, Brilot AF, Pourmal S, Trenker R, Pospiech T, Gupta S, Barsi-Rhyne B, Belyy V, Barile-Hill AW, Nock S, Liu Y, Krogan NJ, Ralston CY, Swaney DL, García-Sastre A, Ott M, Vignuzzi M, Walter P, and Manglik A
- Subjects
- Angiotensin-Converting Enzyme 2 chemistry, Angiotensin-Converting Enzyme 2 immunology, Animals, Antibodies, Neutralizing chemistry, Antibodies, Viral chemistry, Antibody Affinity, Chlorocebus aethiops, Cryoelectron Microscopy, Humans, Neutralization Tests, Protein Binding, Protein Stability, Single-Domain Antibodies chemistry, Spike Glycoprotein, Coronavirus chemistry, Vero Cells, Antibodies, Neutralizing immunology, Antibodies, Viral immunology, Single-Domain Antibodies immunology, Spike Glycoprotein, Coronavirus immunology
- Abstract
The severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) virus enters host cells via an interaction between its Spike protein and the host cell receptor angiotensin-converting enzyme 2 (ACE2). By screening a yeast surface-displayed library of synthetic nanobody sequences, we developed nanobodies that disrupt the interaction between Spike and ACE2. Cryo-electron microscopy (cryo-EM) revealed that one nanobody, Nb6, binds Spike in a fully inactive conformation with its receptor binding domains locked into their inaccessible down state, incapable of binding ACE2. Affinity maturation and structure-guided design of multivalency yielded a trivalent nanobody, mNb6-tri, with femtomolar affinity for Spike and picomolar neutralization of SARS-CoV-2 infection. mNb6-tri retains function after aerosolization, lyophilization, and heat treatment, which enables aerosol-mediated delivery of this potent neutralizer directly to the airway epithelia., (Copyright © 2020 The Authors, some rights reserved; exclusive licensee American Association for the Advancement of Science. No claim to original U.S. Government Works.)
- Published
- 2020
- Full Text
- View/download PDF
30. Comparative host-coronavirus protein interaction networks reveal pan-viral disease mechanisms.
- Author
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Gordon DE, Hiatt J, Bouhaddou M, Rezelj VV, Ulferts S, Braberg H, Jureka AS, Obernier K, Guo JZ, Batra J, Kaake RM, Weckstein AR, Owens TW, Gupta M, Pourmal S, Titus EW, Cakir M, Soucheray M, McGregor M, Cakir Z, Jang G, O'Meara MJ, Tummino TA, Zhang Z, Foussard H, Rojc A, Zhou Y, Kuchenov D, Hüttenhain R, Xu J, Eckhardt M, Swaney DL, Fabius JM, Ummadi M, Tutuncuoglu B, Rathore U, Modak M, Haas P, Haas KM, Naing ZZC, Pulido EH, Shi Y, Barrio-Hernandez I, Memon D, Petsalaki E, Dunham A, Marrero MC, Burke D, Koh C, Vallet T, Silvas JA, Azumaya CM, Billesbølle C, Brilot AF, Campbell MG, Diallo A, Dickinson MS, Diwanji D, Herrera N, Hoppe N, Kratochvil HT, Liu Y, Merz GE, Moritz M, Nguyen HC, Nowotny C, Puchades C, Rizo AN, Schulze-Gahmen U, Smith AM, Sun M, Young ID, Zhao J, Asarnow D, Biel J, Bowen A, Braxton JR, Chen J, Chio CM, Chio US, Deshpande I, Doan L, Faust B, Flores S, Jin M, Kim K, Lam VL, Li F, Li J, Li YL, Li Y, Liu X, Lo M, Lopez KE, Melo AA, Moss FR 3rd, Nguyen P, Paulino J, Pawar KI, Peters JK, Pospiech TH Jr, Safari M, Sangwan S, Schaefer K, Thomas PV, Thwin AC, Trenker R, Tse E, Tsui TKM, Wang F, Whitis N, Yu Z, Zhang K, Zhang Y, Zhou F, Saltzberg D, Hodder AJ, Shun-Shion AS, Williams DM, White KM, Rosales R, Kehrer T, Miorin L, Moreno E, Patel AH, Rihn S, Khalid MM, Vallejo-Gracia A, Fozouni P, Simoneau CR, Roth TL, Wu D, Karim MA, Ghoussaini M, Dunham I, Berardi F, Weigang S, Chazal M, Park J, Logue J, McGrath M, Weston S, Haupt R, Hastie CJ, Elliott M, Brown F, Burness KA, Reid E, Dorward M, Johnson C, Wilkinson SG, Geyer A, Giesel DM, Baillie C, Raggett S, Leech H, Toth R, Goodman N, Keough KC, Lind AL, Klesh RJ, Hemphill KR, Carlson-Stevermer J, Oki J, Holden K, Maures T, Pollard KS, Sali A, Agard DA, Cheng Y, Fraser JS, Frost A, Jura N, Kortemme T, Manglik A, Southworth DR, Stroud RM, Alessi DR, Davies P, Frieman MB, Ideker T, Abate C, Jouvenet N, Kochs G, Shoichet B, Ott M, Palmarini M, Shokat KM, García-Sastre A, Rassen JA, Grosse R, Rosenberg OS, Verba KA, Basler CF, Vignuzzi M, Peden AA, Beltrao P, and Krogan NJ
- Subjects
- Conserved Sequence, Coronavirus Nucleocapsid Proteins genetics, Cryoelectron Microscopy, Humans, Mitochondrial Membrane Transport Proteins genetics, Mitochondrial Precursor Protein Import Complex Proteins, Phosphoproteins genetics, Phosphoproteins metabolism, Protein Conformation, COVID-19 metabolism, Coronavirus Nucleocapsid Proteins metabolism, Host Microbial Interactions, Mitochondrial Membrane Transport Proteins metabolism, Protein Interaction Maps, Severe acute respiratory syndrome-related coronavirus metabolism, SARS-CoV-2 metabolism, Severe Acute Respiratory Syndrome metabolism
- Abstract
The COVID-19 pandemic, caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), is a grave threat to public health and the global economy. SARS-CoV-2 is closely related to the more lethal but less transmissible coronaviruses SARS-CoV-1 and Middle East respiratory syndrome coronavirus (MERS-CoV). Here, we have carried out comparative viral-human protein-protein interaction and viral protein localization analyses for all three viruses. Subsequent functional genetic screening identified host factors that functionally impinge on coronavirus proliferation, including Tom70, a mitochondrial chaperone protein that interacts with both SARS-CoV-1 and SARS-CoV-2 ORF9b, an interaction we structurally characterized using cryo-electron microscopy. Combining genetically validated host factors with both COVID-19 patient genetic data and medical billing records identified molecular mechanisms and potential drug treatments that merit further molecular and clinical study., (Copyright © 2020 The Authors, some rights reserved; exclusive licensee American Association for the Advancement of Science. No claim to original U.S. Government Works.)
- Published
- 2020
- Full Text
- View/download PDF
31. Robust Sequence Determinants of α-Synuclein Toxicity in Yeast Implicate Membrane Binding.
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Newberry RW, Arhar T, Costello J, Hartoularos GC, Maxwell AM, Naing ZZC, Pittman M, Reddy NR, Schwarz DMC, Wassarman DR, Wu TS, Barrero D, Caggiano C, Catching A, Cavazos TB, Estes LS, Faust B, Fink EA, Goldman MA, Gomez YK, Gordon MG, Gunsalus LM, Hoppe N, Jaime-Garza M, Johnson MC, Jones MG, Kung AF, Lopez KE, Lumpe J, Martyn C, McCarthy EE, Miller-Vedam LE, Navarro EJ, Palar A, Pellegrino J, Saylor W, Stephens CA, Strickland J, Torosyan H, Wankowicz SA, Wong DR, Wong G, Redding S, Chow ED, DeGrado WF, and Kampmann M
- Subjects
- Amino Acid Sequence, Humans, Mutation, Parkinson Disease metabolism, Protein Conformation, Saccharomyces cerevisiae metabolism, alpha-Synuclein chemistry, alpha-Synuclein genetics, Saccharomyces cerevisiae drug effects, alpha-Synuclein toxicity
- Abstract
Protein conformations are shaped by cellular environments, but how environmental changes alter the conformational landscapes of specific proteins in vivo remains largely uncharacterized, in part due to the challenge of probing protein structures in living cells. Here, we use deep mutational scanning to investigate how a toxic conformation of α-synuclein, a dynamic protein linked to Parkinson's disease, responds to perturbations of cellular proteostasis. In the context of a course for graduate students in the UCSF Integrative Program in Quantitative Biology, we screened a comprehensive library of α-synuclein missense mutants in yeast cells treated with a variety of small molecules that perturb cellular processes linked to α-synuclein biology and pathobiology. We found that the conformation of α-synuclein previously shown to drive yeast toxicity-an extended, membrane-bound helix-is largely unaffected by these chemical perturbations, underscoring the importance of this conformational state as a driver of cellular toxicity. On the other hand, the chemical perturbations have a significant effect on the ability of mutations to suppress α-synuclein toxicity. Moreover, we find that sequence determinants of α-synuclein toxicity are well described by a simple structural model of the membrane-bound helix. This model predicts that α-synuclein penetrates the membrane to constant depth across its length but that membrane affinity decreases toward the C terminus, which is consistent with orthogonal biophysical measurements. Finally, we discuss how parallelized chemical genetics experiments can provide a robust framework for inquiry-based graduate coursework.
- Published
- 2020
- Full Text
- View/download PDF
32. An ultra-potent synthetic nanobody neutralizes SARS-CoV-2 by locking Spike into an inactive conformation.
- Author
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Schoof M, Faust B, Saunders RA, Sangwan S, Rezelj V, Hoppe N, Boone M, Billesbølle CB, Puchades C, Azumaya CM, Kratochvil HT, Zimanyi M, Deshpande I, Liang J, Dickinson S, Nguyen HC, Chio CM, Merz GE, Thompson MC, Diwanji D, Schaefer K, Anand AA, Dobzinski N, Zha BS, Simoneau CR, Leon K, White KM, Chio US, Gupta M, Jin M, Li F, Liu Y, Zhang K, Bulkley D, Sun M, Smith AM, Rizo AN, Moss F, Brilot AF, Pourmal S, Trenker R, Pospiech T, Gupta S, Barsi-Rhyne B, Belyy V, Barile-Hill AW, Nock S, Liu Y, Krogan NJ, Ralston CY, Swaney DL, García-Sastre A, Ott M, Vignuzzi M, Walter P, and Manglik A
- Abstract
Without an effective prophylactic solution, infections from SARS-CoV-2 continue to rise worldwide with devastating health and economic costs. SARS-CoV-2 gains entry into host cells via an interaction between its Spike protein and the host cell receptor angiotensin converting enzyme 2 (ACE2). Disruption of this interaction confers potent neutralization of viral entry, providing an avenue for vaccine design and for therapeutic antibodies. Here, we develop single-domain antibodies (nanobodies) that potently disrupt the interaction between the SARS-CoV-2 Spike and ACE2. By screening a yeast surface-displayed library of synthetic nanobody sequences, we identified a panel of nanobodies that bind to multiple epitopes on Spike and block ACE2 interaction via two distinct mechanisms. Cryogenic electron microscopy (cryo-EM) revealed that one exceptionally stable nanobody, Nb6, binds Spike in a fully inactive conformation with its receptor binding domains (RBDs) locked into their inaccessible down-state, incapable of binding ACE2. Affinity maturation and structure-guided design of multivalency yielded a trivalent nanobody, mNb6-tri, with femtomolar affinity for SARS-CoV-2 Spike and picomolar neutralization of SARS-CoV-2 infection. mNb6-tri retains stability and function after aerosolization, lyophilization, and heat treatment. These properties may enable aerosol-mediated delivery of this potent neutralizer directly to the airway epithelia, promising to yield a widely deployable, patient-friendly prophylactic and/or early infection therapeutic agent to stem the worst pandemic in a century., Competing Interests: Competing Interests M.Schoof, B.Faust, R.A.Saunders, N.Hoppe, P.Walter, and A.Manglik are inventors on a provisional patent describing anti-Spike nanobodies described in this manuscript.
- Published
- 2020
- Full Text
- View/download PDF
33. Bivalent binding of a fully human IgG to the SARS-CoV-2 spike proteins reveals mechanisms of potent neutralization.
- Author
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Wang B, Asarnow D, Lee WH, Huang CW, Faust B, Ng PML, Ngoh EZX, Bohn M, Bulkley D, Pizzorno A, Tan HC, Lee CY, Minhat RA, Terrier O, Soh MK, Teo FJ, Yeap YYC, Hu Y, Seah SGK, Maurer-Stroh S, Renia L, Hanson BJ, Rosa-Calatrava M, Manglik A, Cheng Y, Craik CS, and Wang CI
- Abstract
In vitro antibody selection against pathogens from naïve combinatorial libraries can yield various classes of antigen-specific binders that are distinct from those evolved from natural infection
1-4 . Also, rapid neutralizing antibody discovery can be made possible by a strategy that selects for those interfering with pathogen and host interaction5 . Here we report the discovery of antibodies that neutralize SARS-CoV-2, the virus responsible for the COVID-19 pandemic, from a highly diverse naïve human Fab library. Lead antibody 5A6 blocks the receptor binding domain (RBD) of the viral spike from binding to the host receptor angiotensin converting enzyme 2 (ACE2), neutralizes SARS-CoV-2 infection of Vero E6 cells, and reduces viral replication in reconstituted human nasal and bronchial epithelium models. 5A6 has a high occupancy on the viral surface and exerts its neutralization activity via a bivalent binding mode to the tip of two neighbouring RBDs at the ACE2 interaction interface, one in the "up" and the other in the "down" position, explaining its superior neutralization capacity. Furthermore, 5A6 is insensitive to several spike mutations identified in clinical isolates, including the D614G mutant that has become dominant worldwide. Our results suggest that 5A6 could be an effective prophylactic and therapeutic treatment of COVID-19., Competing Interests: Competing interests: B.W., W.H.L., C.W.H., P.M.L.N., E.Z.X.N., H.C.T., C.Y.L., R.A.M., M.K.S., F.J.T., Y.Y.C.Y., Y.H., and C.I.W. are listed as inventors of a filed patent for all 27 monoclonal antibodies mentioned in this manuscript. The other authors declare that they have no competing interests.- Published
- 2020
- Full Text
- View/download PDF
34. Smoothened stimulation by membrane sterols drives Hedgehog pathway activity.
- Author
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Deshpande I, Liang J, Hedeen D, Roberts KJ, Zhang Y, Ha B, Latorraca NR, Faust B, Dror RO, Beachy PA, Myers BR, and Manglik A
- Subjects
- Animals, Binding Sites, Biosensing Techniques, Catalytic Domain drug effects, Cell Membrane metabolism, Cholesterol chemistry, Cholesterol metabolism, Cholesterol pharmacology, Hedgehog Proteins metabolism, Ligands, Mice, Models, Molecular, Molecular Dynamics Simulation, Patched-1 Receptor antagonists & inhibitors, Patched-1 Receptor metabolism, Protein Conformation, Protein Stability, Single-Chain Antibodies immunology, Smoothened Receptor antagonists & inhibitors, Smoothened Receptor chemistry, Sterols chemistry, Sterols metabolism, Xenopus Proteins chemistry, Cell Membrane chemistry, Hedgehog Proteins agonists, Signal Transduction drug effects, Smoothened Receptor agonists, Smoothened Receptor metabolism, Sterols pharmacology
- Abstract
Hedgehog signalling is fundamental to embryonic development and postnatal tissue regeneration
1 . Aberrant postnatal Hedgehog signalling leads to several malignancies, including basal cell carcinoma and paediatric medulloblastoma2 . Hedgehog proteins bind to and inhibit the transmembrane cholesterol transporter Patched-1 (PTCH1), which permits activation of the seven-transmembrane transducer Smoothened (SMO) via a mechanism that is poorly understood. Here we report the crystal structure of active mouse SMO bound to both the agonist SAG21k and to an intracellular binding nanobody that stabilizes a physiologically relevant active state. Analogous to other G protein-coupled receptors, the activation of SMO is associated with subtle motions in the extracellular domain, and larger intracellular changes. In contrast to recent models3-5 , a cholesterol molecule that is critical for SMO activation is bound deep within the seven-transmembrane pocket. We propose that the inactivation of PTCH1 by Hedgehog allows a transmembrane sterol to access this seven-transmembrane site (potentially through a hydrophobic tunnel), which drives the activation of SMO. These results-combined with signalling studies and molecular dynamics simulations-delineate the structural basis for PTCH1-SMO regulation, and suggest a strategy for overcoming clinical resistance to SMO inhibitors.- Published
- 2019
- Full Text
- View/download PDF
35. Correction: A Web-Based Acceptance-Facilitating Intervention for Identifying Patients' Acceptance, Uptake, and Adherence of Internet- and Mobile-Based Pain Interventions: Randomized Controlled Trial.
- Author
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Lin J, Faust B, Ebert DD, Krämer L, and Baumeister H
- Abstract
[This corrects the article DOI: 10.2196/jmir.9925.]., (©Jiaxi Lin, Bianca Faust, David Daniel Ebert, Lena Krämer, Harald Baumeister. Originally published in the Journal of Medical Internet Research (http://www.jmir.org), 06.02.2019.)
- Published
- 2019
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36. A Web-Based Acceptance-Facilitating Intervention for Identifying Patients' Acceptance, Uptake, and Adherence of Internet- and Mobile-Based Pain Interventions: Randomized Controlled Trial.
- Author
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Lin J, Faust B, Ebert DD, Krämer L, and Baumeister H
- Subjects
- Adult, Female, Humans, Male, Middle Aged, Mobile Applications, Surveys and Questionnaires, Telemedicine, Chronic Pain therapy, Internet trends
- Abstract
Background: Internet- and mobile-based interventions are effective for the treatment of chronic pain. However, little is known about patients' willingness to engage with these types of interventions and how the uptake of such interventions can be improved., Objective: The aim of this study was to identify people's acceptance, uptake, and adherence (primary outcomes) with regard to an internet- and mobile-based intervention for chronic pain and the influence of an information video as an acceptance-facilitating intervention (AFI)., Methods: In this randomized controlled trial with a parallel design, we invited 489 individuals with chronic pain to participate in a Web-based survey assessing the acceptance of internet- and mobile-based interventions with the offer to receive an unguided internet- and mobile-based intervention for chronic pain after completion. Two versions of the Web-based survey (with and without AFI) were randomly sent to two groups: one with AFI (n=245) and one without AFI (n=244). Participants who completed the Web-based survey with or without AFI entered the intervention group or the control group, respectively. In the survey, the individuals' acceptance of pain interventions, measured with a 4-item scale (sum score ranging from 4 to 20), predictors of acceptance, sociodemographic and pain-related variables, and physical and emotional functioning were assessed. Uptake rates (log in to the intervention) and adherence (number of completed modules) to the intervention was assessed 4 months after intervention access. To examine which factors influence acceptance, uptake rate, and adherence in the internet- and mobile-based interventions, we conducted additional exploratory subgroup analyses., Results: In total, 57 (intervention group) and 58 (control group) participants in each group completed the survey and were included in the analyses. The groups did not differ with regard to acceptance, uptake rate, or adherence (P=.64, P=.56, P=.75, respectively). Most participants reported moderate (68/115, 59.1%) to high (36/115, 31.3%) acceptance, with 9.6% (11/115) showing low acceptance (intervention group: mean 13.91, SD 3.47; control group: mean 13.61, SD 3.50). Further, 67% (38/57, intervention group) and 62% (36/58, control group) had logged into the intervention. In both groups, an average of 1.04 (SD 1.51) and 1.14 (SD 1.90) modules were completed, respectively., Conclusions: The informational video was not effective with regard to acceptance, uptake rate, or adherence. Despite the high acceptance, the uptake rate was only moderate and adherence was remarkably low. This study shows that acceptance can be much higher in a sample participating in an internet- and mobile-based intervention efficacy trial than in the target population in routine health care settings. Thus, future research should focus not only on acceptance and uptake facilitating interventions but also on ways to influence adherence. Further research should be conducted within routine health care settings with more representative samples of the target population., Trial Registration: German Clinical Trial Registration DRKS00006183; http://www.drks.de/drks_web/navigate.do ?navigationId=trial.HTML&TRIAL_ID=DRKS00006183 (Archived by WebCite at http://www.webcitation.org/70ebHDhne)., (©Jiaxi Lin, Bianca Faust, David Daniel Ebert, Lena Krämer, Harald Baumeister. Originally published in the Journal of Medical Internet Research (http://www.jmir.org), 21.08.2018.)
- Published
- 2018
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37. Mixed Training Methods: Effects of Combining Resisted Sprints or Plyometrics with Optimum Power Loads on Sprint and Agility Performance in Professional Soccer Players.
- Author
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Loturco I, Kobal R, Kitamura K, Cal Abad CC, Faust B, Almeida L, and Pereira LA
- Abstract
The aim of this study was to compare the effects of two different mixed training programs (optimum power load [OPL] + resisted sprints [RS] and OPL + vertical/horizontal plyometrics [PL]) on neuromuscular performance of elite soccer players during a short-term training preseason. Eighteen male professional soccer players took part in this study. The athletes were pair-matched in two training groups: OPL + RS and OPL + PL. Unloaded and resisted sprinting speeds at 5-, 10-, 20-, and 30-m, change of direction (COD) speed, and performance in the squat jump (SJ), countermovement jump (CMJ), and horizontal jump (HJ) were assessed pre- and post- a 5-week training period. Magnitude based inference with the effect sizes were used for data analysis. A possible increase in the SJ and CMJ heights and a likely increase in the HJ distance were observed in the OPL + PL group. Meaningful improvements were observed in the COD speed test for both training groups comparing pre- and post-measures. In both unloaded and resisted sprints, meaningful decreases were observed in the sprinting times for all distances tested. This study shows that a mixed training approach which comprises exercises and workloads able to produce positive adaptations in different phases of sprinting can be a very effective strategy in professional soccer players. Moreover, the possibility of combining optimum power loads with resisted sprints and plyometrics emerges as a novel and suitable option for coaches and sport scientists, due to the applicability and efficiency of this strength-power training approach.
- Published
- 2017
- Full Text
- View/download PDF
38. Changing focus of symptoms: A rare case report of Munchhausen's syndrome.
- Author
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Hagen M, Faust B, Kunzelmann N, Tektas OY, Kornhuber J, and Müller HHO
- Published
- 2017
- Full Text
- View/download PDF
39. Colorectal surgical site infection reduction strategies.
- Author
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DeHaas D, Aufderheide S, Gano J, Weigandt J, Ries J, and Faust B
- Subjects
- Academic Medical Centers, Colorectal Surgery methods, Female, Humans, Male, Program Development, Program Evaluation, United States, Colorectal Surgery adverse effects, Patient Care Planning organization & administration, Primary Prevention organization & administration, Quality Improvement, Surgical Wound Infection prevention & control
- Abstract
Objective: PeaceHealth Sacred Heart Medical Center at RiverBend is a 379 bed hospital; 15,060 surgical procedures were performed in 2014 with 254 being colorectal surgical cases. Using the ACS NSQIP program the hospital was identified as a high outlier for surgical site infection (SSI) in all cases and in colorectal procedures in our July 2012 semiannual report (SAR)., Methods: A best practice bundle to reduce SSI's and a colorectal enhanced recovery pathway were concurrently initiated, Results: After implementation of these best practice elements, our ACS NSQIP results showed a reduction in our colorectal SSI rate from 17.58% for 2011 to 5.11% (1st decile/low outlier) in the January 2015 SAR. Our SSI rate for all cases were reduced from 4.87% to 1.71% (1st decile/low outlier) during the same time period. A reduction in length of stay for our colorectal surgical patients was also noted., Conclusions: The ACS NSQIP program was essential to identify colorectal SSI as an area of concern and for providing data to drive quality improvement measures., (Copyright © 2016 Elsevier Inc. All rights reserved.)
- Published
- 2016
- Full Text
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40. Structures and functions of mitochondrial ABC transporters.
- Author
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Schaedler TA, Faust B, Shintre CA, Carpenter EP, Srinivasan V, van Veen HW, and Balk J
- Subjects
- ATP-Binding Cassette Transporters classification, ATP-Binding Cassette Transporters genetics, Animals, Crystallography, X-Ray, Humans, Mitochondrial Proteins chemistry, Mitochondrial Proteins genetics, Mutation, Phylogeny, Plant Proteins chemistry, Plant Proteins genetics, Plant Proteins metabolism, Saccharomyces cerevisiae chemistry, Saccharomyces cerevisiae genetics, Saccharomyces cerevisiae metabolism, ATP-Binding Cassette Transporters metabolism, Mitochondria metabolism, Mitochondrial Proteins metabolism
- Abstract
A small number of physiologically important ATP-binding cassette (ABC) transporters are found in mitochondria. Most are half transporters of the B group forming homodimers and their topology suggests they function as exporters. The results of mutant studies point towards involvement in iron cofactor biosynthesis. In particular, ABC subfamily B member 7 (ABCB7) and its homologues in yeast and plants are required for iron-sulfur (Fe-S) cluster biosynthesis outside of the mitochondria, whereas ABCB10 is involved in haem biosynthesis. They also play a role in preventing oxidative stress. Mutations in ABCB6 and ABCB7 have been linked to human disease. Recent crystal structures of yeast Atm1 and human ABCB10 have been key to identifying substrate-binding sites and transport mechanisms. Combined with in vitro and in vivo studies, progress is being made to find the physiological substrates of the different mitochondrial ABC transporters., (© 2015 Authors; published by Portland Press Limited.)
- Published
- 2015
- Full Text
- View/download PDF
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