TY - JOUR
T1 - Structure and dynamics of the active Gs-coupled human secretin receptor
AU - Dong, Maoqing
AU - Deganutti, Giuseppe
AU - Piper, Sarah J.
AU - Liang, Yi Lynn
AU - Khoshouei, Maryam
AU - Belousoff, Matthew J.
AU - Harikumar, Kaleeckal G.
AU - Reynolds, Christopher A.
AU - Glukhova, Alisa
AU - Furness, Sebastian G.B.
AU - Christopoulos, Arthur
AU - Danev, Radostin
AU - Wootten, Denise
AU - Sexton, Patrick M.
AU - Miller, Laurence J.
N1 - Funding Information:
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 “Dimplot” 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 Å and 20° 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).
Funding Information:
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).
Publisher Copyright:
© 2020, The Author(s).
PY - 2020/12/1
Y1 - 2020/12/1
N2 - 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.
AB - 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.
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U2 - 10.1038/s41467-020-17791-4
DO - 10.1038/s41467-020-17791-4
M3 - Article
C2 - 32811827
AN - SCOPUS:85089545766
SN - 2041-1723
VL - 11
JO - Nature Communications
JF - Nature Communications
IS - 1
M1 - 4137
ER -