Protein Arginylation: Methods and Protocols

ISBN-13: 9781493929344 / Angielski / Twarda / 2015 / 149 str.

Anna S. Kashina

Opis

This volume presents a comprehensive overview of all the existing methods on analysing protein arginylation, from 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. This book also includes essays from the founders of the field, who originally discovered arginylation in the early 1960s and brought it to international recognition. Written in the highly successful Methods in Molecular Biology series format, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls.Cutting-edge and thorough, Protein Arginylation: Methods and Protocols would interest the emerging body of scientists involved in the studies of posttranslational arginylation and the rapidly growing community of researchers working on a broad range of posttranslational modifications, the analysis of which often meets similar challenges and utilizes similar principles as posttranslational arginylation.

Spis treści

1. Protein Arginylation: Over 50 Years 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 150k xg Supernatants of Axoplasm and Injured Vertebrate Nerves
Nicholas A. Ingoglia

5. Preparation of ATE1 Enzyme from Native Mammalian Tissues
Anna Kashina (Adapted from the original article by R. Soffer in consultation with R. Soffer, H. Kaji and A. Kaji)

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. Bacterial Expression and Purification of Recombinant Arginyltransferase (ATE1) and Arg-tRNA Synthetase (RRS) for Arginylation Assays
Junling Wang and Anna Kashina

10. Assaying ATE1 Activity In Vitro
Junling Wang and Anna Kashina

11. High Throughput Arginylation Assay in Microplate Format
Sougata Saha, Junling Wang, and Anna Kashina

12. Assay of Arginyltransferase Activity by a Fluorescent HPLC Method
Koichi Takao

13. Identification of Arginylated Proteins by Mass Spectrometry
Anna Kashina and John R. Yates, III

14. Analysis of Arginylated Peptides by Subtractive Edman Degradation
Anna Kashina and John R. Yates, III

15. 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

16. Applying Arginylation for Bottom-Up Proteomics
H. Alexander Ebhardt 

17. Development of New Tools for the Studies of Protein Arginylation
Anna Kashina