Cargando…

Submolecular probing of the complement C5a receptor–ligand binding reveals a cooperative two-site binding mechanism

A current challenge to produce effective therapeutics is to accurately determine the location of the ligand-biding site and to characterize its properties. So far, the mechanisms underlying the functional activation of cell surface receptors by ligands with a complex binding mechanism remain poorly...

Descripción completa

Detalles Bibliográficos
Autores principales: Dumitru, Andra C., Deepak, R. N. V. Krishna, Liu, Heng, Koehler, Melanie, Zhang, Cheng, Fan, Hao, Alsteens, David
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Nature Publishing Group UK 2020
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7749166/
https://www.ncbi.nlm.nih.gov/pubmed/33339958
http://dx.doi.org/10.1038/s42003-020-01518-8
_version_ 1783625258326032384
author Dumitru, Andra C.
Deepak, R. N. V. Krishna
Liu, Heng
Koehler, Melanie
Zhang, Cheng
Fan, Hao
Alsteens, David
author_facet Dumitru, Andra C.
Deepak, R. N. V. Krishna
Liu, Heng
Koehler, Melanie
Zhang, Cheng
Fan, Hao
Alsteens, David
author_sort Dumitru, Andra C.
collection PubMed
description A current challenge to produce effective therapeutics is to accurately determine the location of the ligand-biding site and to characterize its properties. So far, the mechanisms underlying the functional activation of cell surface receptors by ligands with a complex binding mechanism remain poorly understood due to a lack of suitable nanoscopic methods to study them in their native environment. Here, we elucidated the ligand-binding mechanism of the human G protein-coupled C5a receptor (C5aR). We discovered for the first time a cooperativity between the two orthosteric binding sites. We found that the N-terminus C5aR serves as a kinetic trap, while the transmembrane domain acts as the functional site and both contributes to the overall high-affinity interaction. In particular, Asp282 plays a key role in ligand binding thermodynamics, as revealed by atomic force microscopy and steered molecular dynamics simulation. Our findings provide a new structural basis for the functional and mechanistic understanding of the GPCR family that binds large macromolecular ligands.
format Online
Article
Text
id pubmed-7749166
institution National Center for Biotechnology Information
language English
publishDate 2020
publisher Nature Publishing Group UK
record_format MEDLINE/PubMed
spelling pubmed-77491662020-12-21 Submolecular probing of the complement C5a receptor–ligand binding reveals a cooperative two-site binding mechanism Dumitru, Andra C. Deepak, R. N. V. Krishna Liu, Heng Koehler, Melanie Zhang, Cheng Fan, Hao Alsteens, David Commun Biol Article A current challenge to produce effective therapeutics is to accurately determine the location of the ligand-biding site and to characterize its properties. So far, the mechanisms underlying the functional activation of cell surface receptors by ligands with a complex binding mechanism remain poorly understood due to a lack of suitable nanoscopic methods to study them in their native environment. Here, we elucidated the ligand-binding mechanism of the human G protein-coupled C5a receptor (C5aR). We discovered for the first time a cooperativity between the two orthosteric binding sites. We found that the N-terminus C5aR serves as a kinetic trap, while the transmembrane domain acts as the functional site and both contributes to the overall high-affinity interaction. In particular, Asp282 plays a key role in ligand binding thermodynamics, as revealed by atomic force microscopy and steered molecular dynamics simulation. Our findings provide a new structural basis for the functional and mechanistic understanding of the GPCR family that binds large macromolecular ligands. Nature Publishing Group UK 2020-12-18 /pmc/articles/PMC7749166/ /pubmed/33339958 http://dx.doi.org/10.1038/s42003-020-01518-8 Text en © The Author(s) 2020, corrected publication 2021 https://creativecommons.org/licenses/by/4.0/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/ (https://creativecommons.org/licenses/by/4.0/) .
spellingShingle Article
Dumitru, Andra C.
Deepak, R. N. V. Krishna
Liu, Heng
Koehler, Melanie
Zhang, Cheng
Fan, Hao
Alsteens, David
Submolecular probing of the complement C5a receptor–ligand binding reveals a cooperative two-site binding mechanism
title Submolecular probing of the complement C5a receptor–ligand binding reveals a cooperative two-site binding mechanism
title_full Submolecular probing of the complement C5a receptor–ligand binding reveals a cooperative two-site binding mechanism
title_fullStr Submolecular probing of the complement C5a receptor–ligand binding reveals a cooperative two-site binding mechanism
title_full_unstemmed Submolecular probing of the complement C5a receptor–ligand binding reveals a cooperative two-site binding mechanism
title_short Submolecular probing of the complement C5a receptor–ligand binding reveals a cooperative two-site binding mechanism
title_sort submolecular probing of the complement c5a receptor–ligand binding reveals a cooperative two-site binding mechanism
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7749166/
https://www.ncbi.nlm.nih.gov/pubmed/33339958
http://dx.doi.org/10.1038/s42003-020-01518-8
work_keys_str_mv AT dumitruandrac submolecularprobingofthecomplementc5areceptorligandbindingrevealsacooperativetwositebindingmechanism
AT deepakrnvkrishna submolecularprobingofthecomplementc5areceptorligandbindingrevealsacooperativetwositebindingmechanism
AT liuheng submolecularprobingofthecomplementc5areceptorligandbindingrevealsacooperativetwositebindingmechanism
AT koehlermelanie submolecularprobingofthecomplementc5areceptorligandbindingrevealsacooperativetwositebindingmechanism
AT zhangcheng submolecularprobingofthecomplementc5areceptorligandbindingrevealsacooperativetwositebindingmechanism
AT fanhao submolecularprobingofthecomplementc5areceptorligandbindingrevealsacooperativetwositebindingmechanism
AT alsteensdavid submolecularprobingofthecomplementc5areceptorligandbindingrevealsacooperativetwositebindingmechanism