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...
Autores principales: | , , , , , , |
---|---|
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 |