Sirtuins: Methods and Protocols (Methods in Molecular Biology)

Featuring a diverse array of model organisms and scientific techniques, Sirtuins: Methods and Protocols collects detailed contributions from experts in the field addressing this vital family of genes. Opening with methods to generate sirtuin biology tools, the book continues by covering methods to identify sirtuin substrates, to measure sirtuin activity, and to study sirtuin biology. 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.  Comprehensive and easy to use, Sirtuins: Methods and Protocols presents detailed protocols for sirtuin research that can be followed directly or modified to investigate new areas of sirtuin biology. Part I: Methods to Generate Sirtuin Biology Tools   1. Introduction: Sirtuins in Aging and Diseases             Leonard Guarente   2. Sirtuins in Yeast: Phenotypes and Tools             Scott Tsuchiyama, Elizabeth Kwan, Weiwei Dang, Antonio Bedalov, and Brian K. Kennedy   3. Studying the C. elegans Sirtuins             Mohan Viswanathan and Heidi A. Tissenbaum   4. Genetic and Biochemical Tools for Investigating Sirtuin Function in Drosophila melanogaster             Jason G. Wood, Rachel Whitaker, and Stephen L. Helfand   5. Generating Mammalian Sirtuin Tools for Protein-Interaction Analysis             Kathleen A. Hershberger, Jonathan Motley, Matthew D. Hirschey, and Kristin A. Anderson   Part II: Methods to Identify Sirtuin Substrates   6. Mass Spectrometry-Based Detection of Protein Acetylation             Yu Li, Jeffrey C. Silva, Mary E. Skinner, and David B. Lombard   7. SILAC-Based Quantification of Sirt1-Responsive Lysine Acetylome             Yue Chen, Gozde Colak, and Yingming Zhao   8. Targeted Quantitation of Acetylated Lysine Peptides by Selected Reaction Monitoring Mass Spectrometry             Matthew J. Rardin, Jason M. Held, and Bradford W. Gibson   9. Identification of Deacetylase Substrates with the Biotin Switch Approach             J. Will Thompson, Alex Robeson, and Joshua L. Andersen   10. Assaying Chromatin Sirtuins             Lei Zhong, Barbara Martinez-Pastor, Dafne M. Silberman, Carlos Sebastian, and Raul Mostoslavsky   Part III: Methods to Measure Sirtuin Activity   11. Measurement of Sirtuin Enzyme Activity Using a Substrate-Agnostic Fluorometric Nicotinamide Assay             Basil P. Hubbard and David A. Sinclair   12. Detecting Sirtuin-Catalyzed Deacylation Reactions Using 32P-Labeled NAD and Thin-Layer Chromatography             Anita Zhu, Xiaoyang Su, and Hening Lin   13. Chemical Acetylation And Deacetylation             Kristofer S. Fritz   14. Accurate Measurement of Nicotinamide Adenine Dinucleotide (NAD+) with High-Performance Liquid Chromatography             Jun Yoshino and Shin-ichiro Imai   15. In Vivo Measurement of the Acetylation State of Sirtuin Substrates as a Proxy for Sirtuin Activity             John Dominy, Pere Puigserver, and Carles Cant   Part IV: Methods to Study Sirtuin Biology   16. Oxygen Flux Analysis to Understand the Biological Function of Sirtuins             Dongning Wang, Michelle F. Green, Eoin McDonnell, and Matthew D. Hirschey   17. The Emerging Links Between Sirtuins and Autophagy             In Hye Lee, Jeanho Yun, and Toren Finkel   18. Methods to Study the Role of Sirtuins in Genome Stability             Paloma Martnez-Redondo and Alejandro Vaquero   19. Circadian Measurements of Sirtuin Biology             Kathryn Moynihan Ramsey, Alison H. Affinati, Clara B. Peek, Biliana Marcheva, Hee-Kyung Hong, and Joseph Bass   20. Utilizing Calorie Restriction to Evaluate the Role of Sirtuins in Healthspan and Lifespan of Mice             Jessica Curtis and Rafael de Cabo