Kashina Anna S. (ed.) Protein Arginylation: Methods and Protocols

2nd edition. — Humana Press, 2023. — 289 p. — (Methods in Molecular Biology 2620). — ISBN 978-1-0716-2941-3.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
Arginyltransfera​se:​ A Personal and Historical Perspective
Arginylation in a Partially Purified Fraction of 150 k xg 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 Posttranslationa​l Arginylation of Proteins in Cultured Cells
Assaying ATE1 Activity in Yeast by β-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 Arginyltransfera​se (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 Arginyltransfera​se Activity and Specificity by Peptide Arrays
Assay of Arginyltransfera​se 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 Arginyltransfera​se (ATE1) Iron-Sulfur Cluster
N-Terminal Arginylation Pull-down Analysis Using the R-Catcher Tool
Identification of Protein Arginylation by Encapsulated N-Terminal Peptide Enrichment Method
Monitoring the Activation of Selective Autophagy via N-Terminal Arginylation
Analyzing the Interaction of Arginylated Proteins and Nt-Arg-Mimicking Chemical Compounds to N-Recognins
Synthesis of Stably Charged Arg-tRNAArg for Structural Analysis
A High-Throughput Colorimetric Microplate Assay for Determination of Plasma Arginase Activity
Development of New Tools for the Studies of Protein Arginylation