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

Thomas, Hannah, Dolton, Garry, Fernandez Cardo, Lucia, Ternette, Nicola, Sewell, Andrew K. ORCID: https://orcid.org/0000-0003-3194-3135 and Schmidlin, Thierry 2026. Protocol for MR1-ligand cross-linking to discover metabolite antigens by proteomics. STAR Protocols 7 (1) , 104399. 10.1016/j.xpro.2026.104399

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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.

Item Type:	Article
Date Type:	Published Online
Status:	Published
Schools:	Schools > Medicine
Publisher:	Elsevier BV
ISSN:	2666-1667
Date of First Compliant Deposit:	9 March 2026
Last Modified:	08 May 2026 14:51
URI:	https://orca.cardiff.ac.uk/id/eprint/185625

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