Slow-binding inhibition of NAD+ glycohydrolase by arabino analogues of beta-NAD.
Modifications at the 2'-position of the nicotinamide-ribosyl moiety influence dramatically the nature of the interactions of the modified beta-NAD+ with calf spleen NAD+ glycohydrolase (EC 3.2.2.6), an enzyme that cleaves the nicotinamide-ribose bound in NAD(P)+. Nicotinamide arabinoside adenine dinucleotide (ara-NAD+) and nicotinamide 2'-deoxy-2'-fluoroarabinoside adenine dinucleotide (araF-NAD+) are not hydrolyzed at measurable rates and are the first documented examples of reversible slow binding inhibitors of this class of enzyme. The kinetic data obtained are consistent with both slow kon and koff rate constants in the formation of an enzyme-inhibitor complex, i.e. the association rate constants are about 10(4) and 10(6) slower than diffusion rates, respectively, for araF-NAD+ and ara-NAD+, and the half-life of the complex is about 3-10 min for both analogues. The kinetic model does not account for a slow turnover of an ADP-ribosyl-enzyme intermediary complex. AraF-NAD+ is one of the most potent inhibitors described for NAD+ glycohydrolase.
Muller-Steffner
H M
HM
Laboratoire de Chimie Bioorganique (Centre National de la Recherche Scientifique, Unité de Recherche Associée 1386), Université Louis Pasteur Strasbourg, Faculté de Pharmacie, Illkirch, France.
Malver
O
O
Hosie
L
L
Oppenheimer
N J
NJ
Schuber
F
F
D000818:Animals; D001089:Arabinose; D002417:Cattle; D007700:Kinetics; D008861:Microsomes / Q000201:enzymology; D008962:Models, Theoretical; D009243:NAD / Q000031:analogs & derivatives* / Q000494:pharmacology*; D009244:NAD+ Nucleosidase / Q000037:antagonists & inhibitors*; D011485:Protein Binding; D013154:Spleen / Q000201:enzymology
D009243:NAD; D001089:Arabinose; D009244:NAD+ Nucleosidase