This second edition provides updated and new chapters that detail comprehensive overview of all the existing methods on analyzing protein arginylation. Chapters guide readers through the early methods utilizing crude protein preparations and whole-cell assays to the latest advanced methods involving recombinant protein techniques, antibodies, high precision mass spectrometry, and chemical probes. Written in the format of the highly successful Methods in Molecular Biology series, each chapter includes an introduction to the topic, lists necessary materials and methods, includes tips on troubleshooting and known pitfalls, and step-by-step, readily reproducible protocols.
Cutting-edge and thorough, Protein Arginylation: Methods and Protocols, Second Edition aims to be of interest to novice researchers involved in posttranslational arginylation research and researchers working on a broad range of posttranslational modifications.
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 ?-gal degradation.- Assaying arginylation activity in cell lysates using a fluorescent reporter.- AssayinlÃ3
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