Year: 2025
Type: article
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 µg 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.
Source: Nature Chemical Biology
Authors: Zongtao Lin, Yixuan Xie, Joanna Gongora, Xingyu Liu, Emily Zahn, Bibhuti Bhusana Palai, Daniel H. Ramirez, Richard M. Searfoss, Francisca N. Vitorino, Rashmi Karki, Geoffrey P. Dann, Chenfeng Zhao, Xian Han, Brittany MacTaggart, Xin Lan, Dechen Fu, Lina Greenberg, Yi Zhang, Kory J. Lavine, Michael J. Greenberg, Dongwen Lv, Anna Kashina, Benjamin A. Garcia
Language: English
Subfield: Spectroscopy
Field: Chemistry
Domain: Physical Sciences
Open Access status: hybrid
Estimated APC paid: $12,690
Funders: National Science Foundation, U.S. Department of Health and Human Services, American Heart Association, University of Texas at San Antonio, Children's Discovery Institute, National Institutes of Health, University of Texas Health Science Center at San Antonio, Washington University School of Medicine in St. Louis, National Institute on Aging, National Heart, Lung, and Blood Institute, National Cancer Institute, National Institute of General Medical Sciences, National Institute of Diabetes and Digestive and Kidney Diseases, National Institute of Neurological Disorders and Stroke, Division of Chemistry, Eunice Kennedy Shriver National Institute of Child Health and Human Development
Awards: G100384009, Mays Cancer Center at UT Health SA, G196867973, G2246396969, G2337220457, Hijacking Post-translational Arginylation for Targeted Protein Degradation, Diabetes Research Center, G2686067895, G2994152315, G3168635882, G3219208679, G3439828402, G3861159085, G3874765150, G3964814647, Regulation of cell migration by nucleotide coding sequence and arginylation, Role of arginylation in prevention of alpha synuclein-driven neurodegeneration, G4228147442, G425333985, A Mass Spectrometry Approach to the Genetic and Epigenetic Mechanisms Controlling Neuronal Identity, G4528017521, G4729907715, G4961167568, G5542770389, G5543653603, G6008123622, G7083453410, G757106693, G8024099583, Arginyl-tRNA beyond translation: mechanism and regulation of protein arginylation, Collaborative Research: MFB: Deciphering the Logic of PTM Crosstalk via Novel Chemical Technology: Histones and Beyond, Regulation of cardiac power output in health and disease, G8427634493, Development of Caspase Cleavage Targeting Chimeras (CACTACs) for Targeted Protein Cleavage, Human heart-on-a-chip to study the immune system in cardiac disease pathogenesis and repair, G9000055313, G9000080425, G9000119205, G9000161902, G9000246895, G9000324485, Specification and Function of Tissue Resident and Recruited Macrophages in Cardiac Remodeling and Heart Failure, Alzheimer's Disease Research Center, Quantitative mass spectrometry for comprehending epigenetic mechanisms in a new underlying neurological developmental disorder, G900808053, G901504010, G920047108
OpenAlex ID: W4413495407