Detecting sirtuin-catalyzed deacylation reactions using 32P-labeled NAD and thin-layer chromatography

Anita Zhu, Xiaoyang Su, Hening Lin

Research output: Chapter in Book/Report/Conference proceedingChapter

3 Scopus citations

Abstract

Sirtuins are a class of enzymes with important functions in regulation aging, metabolism, and genome stability. They were originally known as nicotinamide adenine dinucleotide (NAD)-dependent protein lysine deacetylases. However, recently it has been discovered that certain sirtuins with weak deacetylase activity also hydrolyze novel acyl lysine modifications. These findings indicate that other sirtuins with weak deacetylase activity may also possess novel activities on unknown protein posttranslational modifications. Analytical methods that can help to identify new activity of sirtuins and new acyl lysine modifications are thus needed. Here we describe a sensitive method that uses 32P-labeled NAD and thin-layer chromatography to detect sirtuin-catalyzed deacylation reactions. This method can help to discover new acyl lysine modifications that can be removed by novel sirtuin activities.

Original languageEnglish (US)
Title of host publicationSirtuins
Subtitle of host publicationMethods and Protocols
EditorsMatthew Hirschey
Pages179-189
Number of pages11
DOIs
StatePublished - 2013
Externally publishedYes

Publication series

NameMethods in Molecular Biology
Volume1077
ISSN (Print)1064-3745

All Science Journal Classification (ASJC) codes

  • Molecular Biology
  • Genetics

Keywords

  • ADP-ribose
  • CobB
  • Glutamate dehydrogenase
  • Nicotinamide adenine dinucleotide
  • PfSir2a
  • Sirt5
  • Sirtuin
  • Thin-layer chromatography

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