The T-Cell Receptor Can Bind to the Peptide-Bound Major Histocompatibility Complex and Uncomplexed beta(2)-Microglobulin through Distinct Binding Sites

作者：Merkle Patrick S; Irving Melita; Hongjian Song; Ferber Mathias; Jorgensen Thomas J D; Scholten Kirsten; Luescher Immanuel; Coukos George; Zoete Vincent; Cuendet Michel A; Michielin Olivier^*; Rand Kasper D^*

来源：Biochemistry, 2017, 56(30): 3945-3961.

DOI：10.1021/acs.biochem.7b00385

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 beta(2)-constant domain (C-and F-strand) directly interacts with the unbound MHC light chain, beta(2)-microglobulin (beta(2)m). Surface plasmon resonance measurements corroborated a binding event between TCR and fi,m with a dissociation constant of 167 +/- 20 mu M. We propose a model structure for the TCR beta(2)m 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 beta(2)m on T-cell cytotoxicity, suggesting an alternate role for beta 2m binding. Overall, we show that binding of beta(2)m 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:

• 出版日期2017-8-1