Purification of a nitric oxide-stimulated ADP-ribosylated protein using biotinylated beta-nicotinamide adenine dinucleotide.
ADP-ribosylation, involving the transfer of an ADP-ribose moiety from NAD to proteins, is mediated by several bacterial toxins and endogenous ADP-ribosyltransferases. We report here the synthesis of biotinylated NAD and its use to label and purify biotinyl-ADP-ribosylated proteins. We demonstrate that biotinylated NAD can be used by diphtheria toxin to biotinylate elongation factor 2. Using avidin affinity chromatography, we have purified a protein whose ADP-ribosylation is enhanced by nitric oxide and which has been identified as glyceraldehyde-3-phosphate dehydrogenase.
Zhang
J
J
Department of Neuroscience, Johns Hopkins University School of Medicine, Baltimore, Maryland 21205.
Snyder
S H
SH
D000246:Adenosine Diphosphate Ribose / Q000737:chemistry*; D000818:Animals; D001710:Biotin / Q000737:chemistry*; D001923:Brain Chemistry; D002851:Chromatography, High Pressure Liquid; D004591:Electrophoresis, Polyacrylamide Gel; D009243:NAD / Q000138:chemical synthesis*; D009419:Nerve Tissue Proteins / Q000737:chemistry / Q000302:isolation & purification*; D009569:Nitric Oxide / Q000737:chemistry*; D020652:Peptide Elongation Factor 2; D010445:Peptide Elongation Factors / Q000737:chemistry / Q000302:isolation & purification*; D051381:Rats
D009419:Nerve Tissue Proteins; D020652:Peptide Elongation Factor 2; D010445:Peptide Elongation Factors; D009243:NAD; D000246:Adenosine Diphosphate Ribose; D009569:Nitric Oxide; D001710:Biotin