Impact of HLA-DR antigen binding cleft rigidity on T cell recognition

Christopher Szeto, Joseph I. Bloom, Hannah Sloane, Christian A. Lobos, James Fodor, Dhilshan Jayasinghe, Demetra S. M. Chatzileontiadou, Emma J. Grant, Ashley M. Buckle, Stephanie Gras

Research output: Contribution to journalArticleResearchpeer-review

Abstract

The interaction between T cell receptor (TCR) and peptide (p)-Human Leukocyte Antigen (HLA) complexes is the critical first step in determining T cell responses. X-ray crystallographic studies of pHLA in TCR-bound and free states provide a structural perspective that can help understand T cell activation. These structures represent a static “snapshot”, yet the nature of pHLAs and their interactions with TCRs are highly dynamic. This has been demonstrated for HLA class I molecules with in silico techniques showing that some interactions, thought to stabilise pHLA-I, are only transient and prone to high flexibility. Here, we investigated the dynamics of HLA class II molecules by focusing on three allomorphs (HLA-DR1,-DR11 and-DR15) that are able to present the same epitope and activate CD4+ T cells. A single TCR (F24) has been shown to recognise all three HLA-DR molecules, albeit with different affinities. Using molecular dynamics and crystallographic ensemble refinement, we investigate the molecular basis of these different affinities and uncover hidden roles for HLA polymorphic residues. These polymorphisms were responsible for the widening of the antigen binding cleft and disruption of pHLA-TCR interactions, underpinning the hierarchy of F24 TCR binding affinity, and ultimately T cell activation. We expanded this approach to all available pHLA-DR structures and discovered that all HLA-DR molecules were inherently rigid. Together with in vitro protein stability and peptide affinity measurements, our results suggest that HLA-DR1 possesses inherently high protein stability, and low HLA-DM susceptibility.

Original languageEnglish
Article number7081
Number of pages19
JournalInternational Journal of Molecular Sciences
Volume21
Issue number19
DOIs
Publication statusPublished - 25 Sep 2020

Keywords

  • Ensemble refinement
  • Human leukocyte antigen (HLA)
  • MHC class II
  • Molecular dynamics
  • Peptide binding
  • Protein stability
  • TCR binding

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