Mapping the TCR-Peptide-HLA Interface through Hydrogen-Deuterium Exchange Mass Spectrometry.
- 1Immunocore Limited, 92 Park Drive, Milton Park, AbingdonOX14 4RY, United Kingdom.
- 2Astbury Centre for Structural Molecular Biology, School of Molecular and Cellular Biology, Faculty of Biological Sciences, University of Leeds, LeedsLS2 9JT, United Kingdom.
Abstract
T-cell receptor (TCR)-based therapeutics recognize specific peptides presented by human leukocyte antigens (HLAs) on antigen-presenting cells. The docking orientation of TCRs to peptide-HLA (pHLA) complexes is critical for T-cell activation as it underpins signaling and downstream immune responses. Traditionally, TCR binding geometries with pHLAs have been resolved through X-ray crystallography. Here, we demonstrate that conventional peptide-level hydrogen-deuterium exchange mass spectrometry (HDX-MS) workflows can identify productive TCR engagement with their cognate pHLAs. By quantifying differential deuterium uptake upon complex formation, we show that TCRs that bind centrally within the pHLA groove have distinct uptake profiles compared with those adopting alternative orientations. Comparison of uptake data with information from structure prediction further contextualized the HDX data for distinct TCR-pHLA binding modes, thus highlighting the value of integrating experimental and computational approaches. Collectively, our findings establish existing HDX-MS workflows as rapid and accessible approaches for probing TCR-pHLA interactions, with direct implications for the development of TCR-based biotherapeutics.
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BibTeX
@article{powell2026mapping,
title = {{Mapping the TCR-Peptide-HLA Interface through Hydrogen-Deuterium Exchange Mass Spectrometry.}},
author = {Thomas Powell and Alice Colyer and Gemma Wildsmith and Vijaykumar Karuppiah and Saher Afshan Shaikh and Martin Ebner and Andrew Creese and Antonio N Calabrese},
journal = {Analytical chemistry},
year = {2026},
month = sep,
doi = {10.1021/acs.analchem.5c07287},
url = {https://europepmc.org/article/MED/42915246},
note = {CC BY},
}
RIS
TY - JOUR
TI - Mapping the TCR-Peptide-HLA Interface through Hydrogen-Deuterium Exchange Mass Spectrometry.
AU - Thomas Powell
AU - Alice Colyer
AU - Gemma Wildsmith
AU - Vijaykumar Karuppiah
AU - Saher Afshan Shaikh
AU - Martin Ebner
AU - Andrew Creese
AU - Antonio N Calabrese
PY - 2026
DA - 2026/09/01
JO - Analytical chemistry
DO - 10.1021/acs.analchem.5c07287
UR - https://europepmc.org/article/MED/42915246
AB - T-cell receptor (TCR)-based therapeutics recognize specific peptides presented by human leukocyte antigens (HLAs) on antigen-presenting cells. The docking orientation of TCRs to peptide-HLA (pHLA) complexes is critical for T-cell activation as it underpins signaling and downstream immune responses. Traditionally, TCR binding geometries with pHLAs have been resolved through X-ray crystallography. Here, we demonstrate that conventional peptide-level hydrogen-deuterium exchange mass spectrometry (HDX-MS) workflows can identify productive TCR engagement with their cognate pHLAs. By quantifying differential deuterium uptake upon complex formation, we show that TCRs that bind centrally within the pHLA groove have distinct uptake profiles compared with those adopting alternative orientations. Comparison of uptake data with information from structure prediction further contextualized the HDX data for distinct TCR-pHLA binding modes, thus highlighting the value of integrating experimental and computational approaches. Collectively, our findings establish existing HDX-MS workflows as rapid and accessible approaches for probing TCR-pHLA interactions, with direct implications for the development of TCR-based biotherapeutics.
SN - 0003-2700
SN - 1520-6882
ER -