Sir2 products and activities

An analog of 2′-O-acetyl-ADP-ribose or 3′-O-acetyl-ADP-ribose exhibiting increased stability in cells, wherein the 2′-O-acetyl-ADP-ribose and the 3′-O-acetyl-ADP-ribose both comprise an acetate group with an ester oxygen, and an oxygen between two phosphorus atoms.

A method of preparing 2′/3′-O-acetyl-ADP-ribose, the method comprising

• combining a Sir2 enzyme with NAD+ and an acetylated peptide substrate of the Sir2 in a reaction mixture under conditions and for a time sufficient to deacetylate the peptide;
• then
• identifying and purifying the 2′/3′-O-acetyl-ADP-ribose from the reaction mixture.

A method of detecting activity of a Sir2 in a composition, the method comprising

• combining the composition with NAD+ and an acetylated peptide substrate of a Sir2 in a reaction mixture under conditions and for a time sufficient to deacetylate the peptide in the presence of Sir2 activity;
• then measuring 2′/3′-O-acetyl-ADP-ribose;
• wherein the presence of 2′/3′-O-acetyl-ADP-ribose indicates the presence of Sir2 activity in the composition.

A method of deacetylating an acetylated peptide, the method comprising combining the peptide with a Sir2 enzyme and NAD+, wherein the acetylated peptide is not a histone.

A prodrug of 2′/3′-O-acetyl-ADP-ribose, the prodrug comprising 2′/3′-O-acetyl-ADP-ribose covalently bonded to a moiety by an esterase-sensitive bond, wherein the prodrug is capable of passing into a cell more easily than 2′/3′-O-acetyl-ADP-ribose.