Protein Arginylation
Springer-Verlag New York Inc.
978-1-0716-2941-3 (ISBN)
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 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 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 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.
Erscheinungsdatum | 06.04.2023 |
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Reihe/Serie | Methods in Molecular Biology ; 2620 |
Zusatzinfo | 35 Illustrations, color; 21 Illustrations, black and white; X, 297 p. 56 illus., 35 illus. in color. |
Verlagsort | New York, NY |
Sprache | englisch |
Maße | 178 x 254 mm |
Themenwelt | Naturwissenschaften ► Biologie ► Biochemie |
Naturwissenschaften ► Chemie | |
ISBN-10 | 1-0716-2941-7 / 1071629417 |
ISBN-13 | 978-1-0716-2941-3 / 9781071629413 |
Zustand | Neuware |
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