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An Unbiased Proteomic Platform for ATE1-based Arginylation Profiling-Sciex7600

ABSTRACT: Protein arginylation is an essential posttranslational modification (PTM) catalyzed by arginyl-tRNA-protein transferase 1 (ATE1) in mammalian systems. Arginylation features a post-translational conjugation of an arginyl to a protein, making it extremely challenging to differentiate from translational arginine residues with the same mass in a protein sequence. Here we present a general ATE1-based arginylation profiling platform for the unbiased discovery of arginylation substrates and their precise modification sites. This method integrates isotopic arginine labeling into an ATE1 assay utilizing biological lysates (ex vivo) rather than live cells, thus eliminating translational bias derived from the ribosomal activity and enabling bona fide arginylation identification using isotopic features. The method has been successfully applied to an array of peptide, protein, cell, patient, and animal tissue samples using 20 ug sample input, with 235 unique arginylation sites revealed from human proteomes. Representative sites were validated and followed up for their biological functions. The developed platform is globally applicable to the aforementioned sample types and therefore paves the way for functional studies of this difficult-to-characterize protein modification.

INSTRUMENT(S): ZenoTOF 7600

ORGANISM(S): Homo Sapiens (ncbitaxon:9606)

SUBMITTER: Benjamin A. Garcia

PROVIDER: MSV000097197 | MassIVE | Sat Feb 22 20:25:00 GMT 2025

SECONDARY ACCESSION(S): PXD061088

REPOSITORIES: MassIVE

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