Protein Arginylation: Methods and Protocols

ISBN-13: 9781071629413 / Angielski

Anna S. Kashina

Opis

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.

Spis treści

1. Protein arginylation: milestones of discovery

Anna Kashina

2. Recollection of how we came across the protein modification with amino acids by aminoacyl tRNA-protein transferase

Hideko Kaji and Akira Kaji

 

3. Arginyltransferase: a personal and historical perspective

Richard L. Soffer

 

4. Arginylation in a Partially Purified Fraction of 150,000xg Supernatants of Axoplasm and Injured Vertebrate Nerves

Nicholas A. Ingoglia

 

5. Preparation of ATE1 enzyme from native mammalian tissues

Anna Kashina

 

6. Correlated measurement of endogenous ATE1 activity on native acceptor proteins in tissues and cultured cells to detect cellular aging

Hideko Kaji and Akira Kaji

 

7. Assaying the Post-translational Arginylation of Proteins in Cultured Cells

Mauricio R. Galiano and Marta E. Hallak

 

8. Assaying ATE1 activity in yeast by β-gal degradation

Anna Kashina.

 

9. Assaying arginylation activity in cell lysates using a fluorescent reporter

Akhilesh Kumar and Fangliang Zhang.

 

10. Assaying intracellular arginylation activity using a fluorescent reporter. Brittany MacTaggart and Anna Kashina

 

11. Bacterial expression and purification of recombinant arginyltransferase (ATE1) and Arg-tRNA synthetase (RRS) for arginylation assays

 Junling Wang and Anna Kashina

 

12. Preparation of tRNA^Arg for arginylation assay by in vitro transcription by Irem Avcilar-Kucukgoze and Anna Kashina

 

13. Preparation of an enriched tRNA^Arg fraction for arginylation by expression in E. coli

Irem Avcilar-Kucukgoze, Howard Gamper, Jr., Ya-Ming Hou, and Anna Kashina

 

14. Enzymatic aminoacylation of tRNA^Arg using recombinant Arg-tRNA synthetase Irem Avcilar-Kucukgoze and Anna Kashina

 

15. Assaying ATE1 activity in vitro

 Junling Wang and Anna Kashina.

 

16. High throughput arginylation assay in microplate format

Sougata Saha, Junling Wang, and Anna Kashina

 

17. Assaying for arginyltransferase activity and specificity by peptide arrays

Junling Wang and Anna Kashina.

 

18. Assay of arginyltransferase activity by a fluorescent HPLC method

 Koichi Takao

 

19. Identification of arginylated proteins by mass spectrometry

Anna Kashina and John R. Yates, III

 

20. Analysis of arginylated peptides by subtractive Edman degradation. Anna Kashina and John R. Yates, III

21. Transferase-Mediated Labeling of Protein N-Termini with Click Chemistry Handles

Anne M. Wagner, John B. Warner, Haviva E. Garrett, Christopher R. Walters, and E. James Petersson.

 

22. Synthesis of peptides and proteins with site-specific glutamate arginylation

Marie Shimogawa, Yun Huang, Buyan Pan, and E. James Petersson.

 

23. Reconstitution of the arginyltransferase (ATE1) iron-sulfur cluster

Verna Van and Aaron T. Smith.

 

24. N-terminal arginylation pulldown analysis using the R-catcher tool

Taewook Seo, Goeun Han, and Hyunjoo Cha-Molstad

 

25. Identification of protein arginylation by encapsulated N-terminal peptide enrichment method

 Shinyeong Ju and Cheolju Lee.

 

26. Monitoring the Activation of Selective Autophagy via N-terminal Arginylation. Chang Hoon Ji, Su Bin Kim, Min Ju Lee, and Yong Tae Kwon

 

27. Analyzing the Interaction of Arginylated Proteins and Nt-Arg-Mimicking Chemical Compounds to N-Recognins

Chang Hoon Ji, Ah Jung Heo, and Yong Tae Kwon

28 Synthesis of Stably Charged Arg-tRNAArg for Structural Analysis

Yuka Yamaki, Howard Gamper, and Ya-Ming Hou

 

29. A High-Throughput Colorimetric Microplate Assay for Determination of Plasma Arginase Activity

Natalie J Smith, Mahnaz Maddahfar, Bavani Gunasegaran

 Helen M McGuire, and Barbara Fazekas de St Groth.

 

30 Development of new tools for the studies of protein arginylation

Anna Kashina