Protocol for MR1-ligand cross-linking to discover metabolite antigens by proteomics

Thumbnail Image

Files

protocol_for_mr1ligand_crossl-2026080409223454688.pdf (9.29 MB)

Date issued

2026

Authors

Editors

Journal Title

Journal ISSN

Volume Title

Publisher

Reuse License

CC-BY-4.0
Description of rights: CC-BY-4.0
Item type: Item , ZeitschriftenaufsatzAccess status: Open Access ,

Abstract

Here, we present a protocol for the discovery of MR1-bound metabolite ligands by stabilizing Schiff base interactions between MR1 and its ligands. We describe steps for enriching MR1-ligand complexes using MR1-expressing A549 cells and covalently cross-linking ligands to the MR1 lysine 43 residue via reductive amination. We then detail procedures for identifying ligand-modified MR1 peptides by mass spectrometry and for further validation of candidate ligands using MR1 surface upregulation and TCR functional assays.

DOI

https://doi.org/10.25358/openscience-16026

Description

Keywords

Citation

URI

https://openscience.ub.uni-mainz.de/handle/20.500.12030/16047

Published in

STAR Protocols, 7, 1, Cell Press, Cambridge, Mass., 2026, https://doi.org/10.1016/j.xpro.2026.104399

Relationships

Collections

DFG-491381577-G

Endorsement

Review

Supplemented By

Referenced By