Abstract
The class B secretin GPCR (SecR) has broad physiological effects, with target potential for treatment of metabolic and cardiovascular disease. Molecular understanding of SecR binding and activation is important for its therapeutic exploitation. We combined cryo-electron microscopy, molecular dynamics, and biochemical cross-linking to determine a 2.3 Å structure, and interrogate dynamics, of secretin bound to the SecR:Gs complex. SecR exhibited a unique organization of its extracellular domain (ECD) relative to its 7-transmembrane (TM) core, forming more extended interactions than other family members. Numerous polar interactions formed between secretin and the receptor extracellular loops (ECLs) and TM helices. Cysteine-cross-linking, cryo-electron microscopy multivariate analysis and molecular dynamics simulations revealed that interactions between peptide and receptor were dynamic, and suggested a model for initial peptide engagement where early interactions between the far N-terminus of the peptide and SecR ECL2 likely occur following initial binding of the peptide C-terminus to the ECD.
Original language | English |
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Article number | 4137 |
Number of pages | 17 |
Journal | Nature Communications |
Volume | 11 |
DOIs | |
Publication status | Published - 18 Aug 2020 |
Bibliographical note
Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/ licenses/by/4.0/.Funder
The work was supported by a grant from the National Institutes of Health, GM-132095 (L.J.M. and P.M.S.), the Monash University Ramaciotti Centre for Cryo-Electron Microscopy, the Monash MASSIVE high-performance computing facility, the National Health and Medical Research Council of Australia (NHMRC) project grant (1120919), and NHMRC program grant (1150083). P.M.S. and A.C. are NHMRC Senior Principal Research Fellows and D.W. is an NHMRC Senior Research Fellow. C.A.R. holds a Royal Society Industry Fellowship and acknowledges a grant from the United Kingdom Biotechnology and Biological Sciences Research Council (BB/M006883/1). R.D. was supported by Takeda Science Foundation 2019 Medical Research Grant and Japan Science and Technology Agency PRESTO (18069571).Funding
The work was supported by a grant from the National Institutes of Health, GM-132095 (L.J.M. and P.M.S.), the Monash University Ramaciotti Centre for Cryo-Electron Microscopy, the Monash MASSIVE high-performance computing facility, the National Health and Medical Research Council of Australia (NHMRC) project grant (1120919), and NHMRC program grant (1150083). P.M.S. and A.C. are NHMRC Senior Principal Research Fellows and D.W. is an NHMRC Senior Research Fellow. C.A.R. holds a Royal Society Industry Fellowship and acknowledges a grant from the United Kingdom Biotechnology and Biological Sciences Research Council (BB/M006883/1). R.D. was supported by Takeda Science Foundation 2019 Medical Research Grant and Japan Science and Technology Agency PRESTO (18069571). Model residue interaction analysis. Interactions in the PDB (6WZG), between the chains of the peptide and receptor (P:R) or receptor and G proteins (R:A and R:B), were analyzed using the \u201CDimplot\u201D module within the Ligplot+ program (v2.2)44. Hydrogen bonds were additionally analyzed using the UCSF ChimeraX package, with relaxed distance and angle criteria (0.4 A\u030A and 20\u00B0 tolerance, respectively). Additional analyses and production of images were performed using the UCSF Chimera package (v1.14) from the Computer Graphics Laboratory, University of California, San Francisco (supported by NIH P41 RR-01081) and ChimeraX45 (support from National Institutes of Health R01-GM129325).
Funders | Funder number |
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Biotechnology and Biological Sciences Research Council | BB/M006883/1 |
National Institutes of Health | R01GM132095 |
Takeda Science Foundation | |
National Health and Medical Research Council | 1150083, 1120919 |
Japan Science and Technology Agency | 18069571 |
Keywords
- Cryoelectron microscopy
- Gastroenterology
- Structural biology
ASJC Scopus subject areas
- General Chemistry
- General Biochemistry,Genetics and Molecular Biology
- General Physics and Astronomy
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Giuseppe Deganutti
- Centre for Health and Life Sciences - Assistant Professor (Research)
Person: Teaching and Research
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Chris Reynolds
- Centre for Health and Life Sciences - Professor -Therapeutics and Disease Prevention / Theme Lead
Person: Teaching and Research