Enzymatically Recombinant cIII Deacytylases

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Reversible acetylation of histone and nonhistone proteins is emerging as a major mechanism for regulating protein function.

Acetylation is a pervasive modification involved in most biological functions.

Class III Histone Deacetylases (HDACs) have generally proven easier to manipulate than class I and II HDACs, hence are being employed in the present review.

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Preparation of enzymatically active recombinant
class III protein deacetylases
Brian J.North , Bjoern Schwer , Nidhi Ahuja , Brett Marshall, Eric Verdin ¤

Reversible acetylation of histone and nonhistone proteins is emerging as a major mechanism for regulating protein function.

Acetylation is a pervasive modification involved in most biological functions.

Class III Histone Deacetylases (HDACs) have generally proven easier to manipulate than class I and II HDACs, hence are being employed in the present review.
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.

There are three classes of HDACs, which include eighteen distinct human protein deacetylases, which are grouped on the basis of their primary homology to three protein deacetylases in Saccharomyces cerevisiae.

•Class I histone deacetylases
•Class II histone deacetylases
•Class III histone deacetylases

Class I Histone Deacetylases

•These are homologous to yRPD3, sharing a compact structure.
•The class includes HDAC1, -2, -3, ...