Journal
BIOCHEMISTRY
Volume 56, Issue 30, Pages 3945-3961Publisher
AMER CHEMICAL SOC
DOI: 10.1021/acs.biochem.7b00385
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Funding
- Danish Council for Independent Research\Natural Sciences [11-104058]
- Marie Curie Actions Programme of the European Union [PCIG09-GA-2011-294214]
- Human Frontiers Science Programme (HFSP)
- Villum Fonden [00007292] Funding Source: researchfish
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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:
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