Protein Arginylation

Protein arginylation: milestones of discovery.- Recollection of how we came across the protein modification with amino acids by aminoacyl tRNA-protein transferase.- Arginyltransferase: a personal and historical perspective.- Arginylation in a Partially Purified Fraction of 150,000xg Supernatants of Axoplasm and Injured Vertebrate Nerves.- Preparation of ATE1 enzyme from native mammalian tissues.- Correlated measurement of endogenous ATE1 activity on native acceptor proteins in tissues and cultured cells to detect cellular aging.- Assaying the Post-translational Arginylation of Proteins in Cultured Cells.- Assaying ATE1 activity in yeast by beta-gal degradation.- Assaying arginylation activity in cell lysates using a fluorescent reporter.- Assaying intracellular arginylation activity using a fluorescent reporter.- Bacterial expression and purification of recombinant arginyltransferase (ATE1) and Arg-tRNA synthetase (RRS) for arginylation assays.- Preparation of tRNAArg for arginylation assay by in vitro transcription.- Preparation of an enriched tRNAArg fraction for arginylation by expression in E. coli.- Enzymatic aminoacylation of tRNAArg using recombinant Arg-tRNA synthetase.- Assaying ATE1 activity in vitro.- High throughput arginylation assay in microplate format.- Assaying for arginyltransferase activity and specificity by peptide arrays.- Assay of arginyltransferase activity by a fluorescent HPLC method.- Identification of arginylated proteins by mass spectrometry.- Analysis of arginylated peptides by subtractive Edman degradation.- Transferase-Mediated Labeling of Protein N-Termini with Click Chemistry Handles.- Synthesis of peptides and proteins with site-specific glutamate arginylation.- Reconstitution of the arginyltransferase (ATE1) iron-sulfur cluster.- N-terminal arginylation pulldown analysis using the R-catcher tool.- Identification of protein arginylation by encapsulated N-terminal peptide enrichmentmethod.- Monitoring the