Protein Acetylation: Methods and Protocols

Description

The book focuses on the regulation of protein function and biological activity by the post-translational modification known as acetylation. Chapters detail a wide range of topics in protein acetylation dealing with pathways relevant to cellular homeostasis, gene expression, and DNA metabolism. 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. Authoritative and cutting-edge, Protein Acetylation: Methods and Protocols aims to be an extremely valuable resource to novices and experts alike whose research takes them in the direction of characterizing regulatory switches in protein acetylation events, and how they are manifested in significant ways in diverse mechanisms biological systems. Front Matter ....Pages i-xiii Front Matter ....Pages 1-1 High-Resolution Mass Spectrometry to Identify and Quantify Acetylation Protein Targets (Birgit Schilling, Jesse G. Meyer, Lei Wei, Melanie Ott, Eric Verdin)....Pages 3-16 Isolation of Proteins on Nascent Chromatin and Characterization by Quantitative Mass Spectrometry (Paula A. Agudelo Garcia, Miranda Gardner, Michael A. Freitas, Mark R. Parthun)....Pages 17-27 Detection and Quantitation of Acetylated Histones on Replicating DNA Using In Situ Proximity Ligation Assay and Click-It Chemistry (Pavlo Lazarchuk, Sunetra Roy, Katharina Schlacher, Julia Sidorova)....Pages 29-45 Front Matter ....Pages 47-47 Quantification of In Vitro Protein Lysine Acetylation by Reversed Phase HPLC (Catherine W. Njeri, Onyekachi E. Ononye, Lata Balakrishnan)....Pages 49-56 Measurement and Analysis of Lysine Acetylation by KAT Complexes In Vitro and In Vivo (Anahita Lashgari, Jean-Philippe Lambert, Jacques Côté)....Pages 57-77 Site-Specific Lysine Acetylation Stoichiometry Across Subcellular Compartments (Anastasia J. Lindahl, Alexis J. Lawton, Josue Baeza, James A. Dowell, John M. Denu)....Pages 79-106 Lysine Acetylation of Proteins and Its Characterization in Human Systems (David K. Orren, Amrita Machwe)....Pages 107-130 Front Matter ....Pages 131-131 Molecular and Cellular Characterization of SIRT1 Allosteric Activators (Michael B. Schultz, Conrad Rinaldi, Yuancheng Lu, João A. Amorim, David A. Sinclair)....Pages 133-149 Assessment of SIRT2 Inhibitors in Mouse Models of Cancer (Yashira L. Negrón Abril, Irma Fernández, Robert S. Weiss)....Pages 151-171 Front Matter ....Pages 173-173 Using Yeast as a Model Organism to Study the Functional Roles of Histone Acetylation in DNA Excision Repair (Amelia J. Hodges, Steven A. Roberts, John J. Wyrick)....Pages 175-190 Biochemical and Cellular Assays to Assess the Effects of Acetylation on Base Excision Repair Enzymes (Shrabasti Roychoudhury, Suravi Pramanik, Hannah L. Harris, Kishor K. Bhakat)....Pages 191-206 Analysis of DNA Processing Enzyme FEN1 and Its Regulation by Protein Lysine Acetylation (Onyekachi E. Ononye, Catherine W. Njeri, Lata Balakrishnan)....Pages 207-224 Examining the Role of HDACs in DNA Double-Strand Break Repair in Neurons (Ping-Chieh Pao, Jay Penney, Li-Huei Tsai)....Pages 225-234 Front Matter ....Pages 235-235 Experimental Approaches to Investigate the Role of Helicase Acetylation in Regulating R-Loop Stability (Chenlin Song, Ingrid Grummt)....Pages 237-253 Rapid Detection of p53 Acetylation Status in Response to Cellular Stress Signaling (Marina Farkas, Steven B. McMahon)....Pages 255-262 Front Matter ....Pages 263-263 Isolation and Quantification Brain Region-Specific and Cell Subtype-Specific Histone (De)Acetylation in Cognitive Neuroepigenetics (Craig Myrum, Peter R. Rapp)....Pages 265-277 Functional Analysis of HDACs in Tumorigenesis (Melissa Hadley, Satish Noonepalle, Debarati Banik, Alejandro Villagra)....Pages 279-307 Back Matter ....Pages 309-310