Sirtuins: Methods and Protocols

ISBN-13: 9781627036368 / Angielski / Twarda / 2013 / 314 str.

Matthew D. Hirschey

Opis

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.

Spis treści

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 Cantó

 

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 Martínez-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