| Title: | Chemical modification of aldehyde dehydrogenase by a vinyl ketone analogue of an insect pheromone |
| Author(s): | Blatter EE; Tasayco ML; Prestwich G; Pietruszko R; |
| Address: | "Center of Alcohol Studies, Rutgers University, Piscataway, NJ 08855-0969" |
| ISSN/ISBN: | 0264-6021 (Print) 1470-8728 (Electronic) 0264-6021 (Linking) |
| Abstract: | "A major component of the sex pheromone from the tobacco budworm moth Heliothis virescens is a C16 straight-chain aldehyde with a single unsaturation at the eleventh position. The sex pheromones are inactivated when metabolized to their corresponding acids by insect aldehyde dehydrogenase. During this investigation it was demonstrated that the C16 aldehyde is a good substrate for human aldehyde dehydrogenase (EC 1.2.1.3) isoenzymes E1 and E2 with Km and Kcat. values at pH 7.0 of 2 microM and 0.4 mumol of NADH/min per mg and of 0.6 microM and 0.24 mumol of NADH/min per mg respectively. A vinyl ketone analogue of the pheromone inhibited insect pheromone metabolism; it also inactivated human aldehyde dehydrogenase. Total inactivation of both isoenzymes was achieved at stoichiometric (equal or less than the subunit number) concentrations of vinyl ketone, incorporating 2.1-2.6 molecules/molecule of enzyme. Substrate protection was observed in the presence of the parent aldehyde and 5'-AMP. Peptide maps of tryptic digests of the E2 isoenzyme modified with 3H-labelled vinyl ketone showed that incorporation occurred into a single peptide peak. The labelled peptide of E2 isoenzyme was further purified on h.p.l.c. and sequenced. The label was incorporated into cysteine-302 in the primary structure of E2 isoenzyme, thus indicating that cysteine-302 is located in the aldehyde substrate area of the active site of aldehyde dehydrogenase. Affinity labelling of aldehyde dehydrogenase with vinyl ketones may prove to be of general utility in biochemical studies of these enzymes" |
| Keywords: | Aldehyde Dehydrogenase/*metabolism Aldehydes/chemical synthesis/*pharmacology Alkadienes/chemical synthesis/*pharmacology Amino Acid Sequence Animals Humans Indicators and Reagents Isoenzymes/*metabolism Kinetics Liver/enzymology Molecular Sequence Data M; |
| Notes: | "MedlineBlatter, E E Tasayco, M L Prestwich, G Pietruszko, R eng KO5 AA00046/AA/NIAAA NIH HHS/ R0I AA00186/AA/NIAAA NIH HHS/ Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, Non-P.H.S. Research Support, U.S. Gov't, P.H.S. England 1990/12/01 Biochem J. 1990 Dec 1; 272(2):351-8. doi: 10.1042/bj2720351" |
