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E-raamat: Protein Arginylation: Methods and Protocols

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Protein Arginylation: Methods and ProtocolsSuurem pilt
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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.- 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.
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