Intrinsic alcohol dehydrogenase and hydroxysteroid dehydrogenase activities of human mitochondrial short-chain L-3-hydroxyacyl-CoA dehydrogenase

• Published in: Biochemical journal 2000-01, Vol.345 Pt 1 (1), p.139-143
• Main Authors: He, X Y, Yang, Y Z, Schulz, H, Yang, S Y
• Format: Article
• Language: English
• Subjects: 3-Hydroxyacyl CoA Dehydrogenases - genetics; 3-Hydroxyacyl CoA Dehydrogenases - metabolism; Alcohol Dehydrogenase - genetics; Alcohol Dehydrogenase - metabolism; Alzheimer Disease - enzymology; Alzheimer Disease - etiology; Brain - enzymology; Carrier Proteins - genetics; Carrier Proteins - metabolism; Humans; Hydroxysteroid Dehydrogenases - genetics; Hydroxysteroid Dehydrogenases - metabolism; In Vitro Techniques; Kinetics; Mitochondria - enzymology; Mutation; Recombinant Proteins - genetics; Recombinant Proteins - metabolism; Substrate Specificity
• ISSN: 0264-6021; 1470-8728
• DOI: 10.1042/0264-6021:3450139

The alcohol dehydrogenase (ADH) activity of human short-chain l-3-hydroxyacyl-CoA dehydrogenase (SCHAD) has been characterized kinetically. The k(cat) of the purified enzyme was estimated to be 2. 2 min(-1), with apparent K(m) values of 280 mM and 22microM for 2-propanol and NAD(+), respectively. The k(cat) of the ADH activity was three orders of magnitude less than the l-3-hydroxyacyl-CoA dehydrogenase activity but was comparable with that of the enzyme's hydroxysteroid dehydrogenase (HSD) activity for oxidizing 17beta-oestradiol [He, Merz, Mehta, Schulz and Yang (1999) J. Biol. Chem. 274, 15014-15019]. However, the k(cat) values of intrinsic ADH and HSD activities of human SCHAD were found to be two orders of magnitude less than those reported for endoplasmic-reticulum-associated amyloid beta-peptide-binding protein (ERAB) [Yan, Shi, Zhu, Fu, Zhu, Zhu, Gibson, Stern, Collison, Al-Mohanna et al. (1999) J. Biol. Chem. 274, 2145-2156]. Since human SCHAD and ERAB apparently possess identical amino acid sequences, their catalytic properties should be identical. The recombinant SCHAD has been confirmed to be the right gene product and not a mutant variant. Steady-state kinetic measurements and quantitative analyses reveal that assay conditions such as pH and concentrations of coenzyme and substrate do not account for the kinetic differences reported for ERAB and SCHAD. Rather problematic experimental procedures appear to be responsible for the unrealistically high catalytic rate constants of ERAB. Eliminating the confusion surrounding the catalytic properties of this important multifunctional enzyme paves the way for exploring its role(s) in the pathogenesis of Alzheimer's disease.