Role of NAD(+) in the deacetylase activity of the SIR2-like proteins

Biochem Biophys Res Commun. 2000 Nov 30;278(3):685-90. doi: 10.1006/bbrc.2000.3854.


In this report we describe the role of NAD(+) in the deacetylation reaction catalyzed by the SIR2 family of enzymes. We first show that the products of the reaction detected by HPLC analysis are ADP-ribose, nicotinamide, and a deacetylated peptide substrate. These products are in a 1:1:1 molar ratio, indicating that deacetylation involves the hydrolysis of one NAD(+) to ADP-ribose and nicotinamide for each acetyl group removed. Three results suggest that deacetylation requires an enzyme-ADP-ribose intermediate. First, the enzyme can promote an NAD(+) if nicotinamide exchange reaction that depends on an acetylated substrate. Second, a non-hydrolyzable NAD(+) analog is a competitive inhibitor of the enzyme, and, third, nicotinamide shows product inhibition of deacetylase activity.

Publication types

  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Adenosine Diphosphate Ribose / metabolism
  • Amino Acid Sequence
  • Chromatography, High Pressure Liquid
  • Histone Deacetylases / metabolism*
  • Histones / chemistry
  • Histones / metabolism
  • Kinetics
  • Molecular Sequence Data
  • NAD / metabolism*
  • Niacinamide / metabolism
  • Peptide Fragments / chemistry
  • Peptide Fragments / metabolism
  • Silent Information Regulator Proteins, Saccharomyces cerevisiae*
  • Sirtuin 2
  • Sirtuins
  • Substrate Specificity
  • Trans-Activators / metabolism*


  • Histones
  • Peptide Fragments
  • Silent Information Regulator Proteins, Saccharomyces cerevisiae
  • Trans-Activators
  • NAD
  • Adenosine Diphosphate Ribose
  • Niacinamide
  • SIR2 protein, S cerevisiae
  • Sirtuin 2
  • Sirtuins
  • Histone Deacetylases