The T-Cell Receptor Can Bind to the Peptide-Bound Major Histocompatibility Complex and Uncomplexed β2-Microglobulin through Distinct Binding Sites.
Détails
ID Serval
serval:BIB_7CF39E44F352
Type
Article: article d'un périodique ou d'un magazine.
Collection
Publications
Institution
Titre
The T-Cell Receptor Can Bind to the Peptide-Bound Major Histocompatibility Complex and Uncomplexed β2-Microglobulin through Distinct Binding Sites.
Périodique
Biochemistry
ISSN
1520-4995 (Electronic)
ISSN-L
0006-2960
Statut éditorial
Publié
Date de publication
2017
Peer-reviewed
Oui
Volume
56
Numéro
30
Pages
3945-3961
Langue
anglais
Résumé
T-Cell receptor (TCR)-mediated recognition of the peptide-bound major histocompatibility complex (pMHC) initiates an adaptive immune response against antigen-presenting target cells. The recognition events take place at the TCR-pMHC interface, and their effects on TCR conformation and dynamics are controversial. Here, we have measured the time-resolved hydrogen/deuterium exchange (HDX) of a soluble TCR in the presence and absence of its cognate pMHC by mass spectrometry to delineate the impact of pMHC binding on solution-phase structural dynamics in the TCR. Our results demonstrate that while TCR-pMHC complex formation significantly stabilizes distinct CDR loops of the TCR, it does not trigger structural changes in receptor segments remote from the binding interface. Intriguingly, our HDX measurements reveal that the TCR α-constant domain (C- and F-strand) directly interacts with the unbound MHC light chain, β2-microglobulin (β2m). Surface plasmon resonance measurements corroborated a binding event between TCR and β2m with a dissociation constant of 167 ± 20 μM. We propose a model structure for the TCR-β2m complex based on a refined protein-protein docking approach driven by HDX data and information from molecular dynamics simulations. Using a biological assay based on TCR gene-engineered primary human T cells, we did not observe a significant effect of β2m on T-cell cytotoxicity, suggesting an alternate role for β2m binding. Overall, we show that binding of β2m to the TCR occurs in vitro and, as such, not only should be considered in structure-function studies of the TCR-pMHC complex but also could play a hitherto unidentified role in T-cell function in vivo.
Mots-clé
Amino Acid Substitution, Binding Sites, Cells, Cultured, Cytotoxicity, Immunologic, Deuterium Exchange Measurement, Humans, Kinetics, Ligands, Major Histocompatibility Complex, Models, Molecular, Molecular Docking Simulation, Molecular Dynamics Simulation, Mutation, Peptide Fragments/chemistry, Peptide Fragments/genetics, Peptide Fragments/metabolism, Protein Conformation, Protein Engineering, Protein Interaction Domains and Motifs, Protein Stability, Receptors, Antigen, T-Cell, alpha-beta/chemistry, Receptors, Antigen, T-Cell, alpha-beta/genetics, Receptors, Antigen, T-Cell, alpha-beta/metabolism, Recombinant Proteins/chemistry, Recombinant Proteins/metabolism, T-Lymphocytes/cytology, T-Lymphocytes/immunology, T-Lymphocytes/metabolism, beta 2-Microglobulin/chemistry, beta 2-Microglobulin/genetics, beta 2-Microglobulin/metabolism
Pubmed
Web of science
Création de la notice
27/07/2017 13:14
Dernière modification de la notice
20/08/2019 14:38