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Methods. 2005 Aug;36(4):338-45.

Preparation of enzymatically active recombinant class III protein deacetylases.

Author information

1
Gladstone Institute of Virology and Immunology, University of California, San Francisco, CA, USA.

Abstract

Class III histone deacetylases, or sirtuins, are homologous to the Saccharomyces cerevisiae transcriptional regulator SIR2. The class III enzymes are characterized by their dependence on nicotinamide adenine dinucleotide (NAD+). This cofactor serves as an acetyl-group acceptor in the deacetylation reaction generating O-acetyl-ADP-ribose. Enzymatic activity of sirtuin can be measured in vitro using recombinant proteins purified from mammalian cells after overexpression or after purification from Escherichia coli. This review discusses protocols for the purification of enzymatically active human sirtuin 1, 2, and 3 and their activities on histone and nonhistone substrates.

PMID:
16091304
DOI:
10.1016/j.ymeth.2005.03.004
[Indexed for MEDLINE]

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