Protein Acetylation: Methods and Protocols

ISBN-13: 9781627033046 / Angielski / Twarda / 2013 / 266 str.

Sandra B. Hake; Christian Janzen

Opis

Thousands of proteins have been identified to be acetylated. Immense research power has been dedicated to experiments to solve the biological implications of each and every protein acetylation. Two particular sites of protein acetylation have been described intensively: the N-terminal methionine residue of a nascent protein and lysine residues within a protein. In Protein Acetylation: Methods and Protocols, expert researchers in the field detail many of the methods which are now commonly used to study protein acetylation. These include methods and techniques for identification of protein acetylation, column- and gel electrophoresis-based approaches, computationally prediction, and the biological response to protein acetylation. 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 key tips on troubleshooting and avoiding known pitfalls.Authoritative and practical, Protein Acetylation: Methods and Protocols seeks to aid scientists in the further study of the technical aspects involved in understanding protein acetylation.

Spis treści

1. Validation of Protein Acetylation by Mass Spectrometry

Barry M. Zee and Benjamin A. Garcia

2. Application of the CIRAD Mass Spectrometry Apprach for Lysine Acetylation Site Discovery

Caroline A Evans, Saw Yen Ow, Duncan L Smith, Bernard M Corfe, and Phillip C Wright

3. Application of the MIDAS Approach for Analysis of Lysine Acetylation Sites

Caroline A Evans, John R Griffiths, Richard D Unwin, Anthony D Whetton, Bernard M Corfe

4. Application of High Content Biology to Yield Quantitative Spatial Proteomic Information on Protein Acetylations

Bernard M Corfe, Josephine Kilner, Joanna Chowdry, Roderick SP Benson, Gareth J Griffiths,  Caroline A Evans

5. Towards the N-terminal Acetylome: An N-terminal Acetylated Peptide Enrichment Method Using CNBr-Activated Sepharose Resin

Xumin Zhang and Peter Højrup

 6. Identification and Analysis of O-acetylated Sialoglycoproteins

Chandan Mandal and Chitra Mandal

 7. HPLC-based Quantification of in vitro N-terminal Acetylation

Rune H. Evjenth, Petra Van Damme, Kris Gevaert, Thomas Arnesen

8. Separation and Purification of Multiply Acetylated Proteins using Cation-exchange Chromatography

Romeo Papazyan and Sean D. Taverna

 9. In-gel N-acetylation for the Quantification of the Degree of Protein in vivo N-terminal Acetylation

Petra Van Damme, Thomas Arnesen, Bart Ruttens, Kris Gevaert

 10. Computational Prediction of Lysine Acetylation Proteome-wide

Amrita Basu

11. Generation and Characterization of Pan-specific Anti-acetyllysine Antibody

Dr. Wei Xu and Dr. Shimin Zhao

12. Using Functional Proteome Microarrarys to Study Protein Lysine Acetylation

Jin-Ying Lu, Yu-yi Lin, Jef D. Boeke, and Heng Zhu

13. Quantitation of Nucleosome Acteylation and other Histone Post-Translational Using Microscale NU-ELISA

Bo Dai, Charles Giardina, Theodore P. Rasmussen

14. Preparing Semisynthetic and Fully Synthetic Histones H3 and H4 to Modify the Nucleosome Core

John C. Shimko, Cecil J. Howard, Michael G. Poirier, and Jennifer J. Ottesen

 

^{15. Production of Amino-terminally Acetylated Recombinant Proteins in E. coli}

^{Matthew Johnson, Michael A. Geeves and Daniel P. Mulvihill}

 

^{16. Identification of Lysine Acetyltransferase Substrates using Bioorthogonal Chemical Proteomics}

^{Markus Grammel and Howard C. Hang}

17. Non-radioactive in-vitro Assays for Histone Deacetylases

Alexander-Thomas Hauser, Julia M. Gajer (née Wagner), and Manfred Jung

 18. The Fluorescence-Based Acetylation Assay Using Thiol-Sensitive Probes

Tielong Gao, Chao Yang, Yujun George Zheng

19. Analysis of Protein Acetyltransferase Structure-function Relation by Surface-Enhanced Raman Scattering (SERS): A Tool to Screen and Characterize Small Molecule Modulators

Mohammed Arif, Dhanasekaran Karthigeyan, Soumik Siddhanta, G. V. Pavan Kumar, Chandrabhas Narayana, and Tapas K. Kundu