Delta 5-3 beta-hydroxysteroid dehydrogenase-isomerase activity in canine pancreas.
Activity of delta 5-3 beta-hydroxysteroid dehydrogenase coupled with steroid-delta 5-4-isomerase was demonstrated for the first time in the pancreas. The enzyme complex was assayed by measuring the conversion of pregnenolone to progesterone as well as of dehydroepiandrosterone to androstenedione and found to be localized primarily in the mitochondrial fraction of dog pancreas homogenates. The delta 5-3 beta-hydroxysteroid dehydrogenase used either NAD+ or NADP+ as co-substrates, although maximal activity was observed with NAD+. In phosphate buffer, pH 7.0 and 37 degrees C, the apparent Km values of the dehydrogenase were 6.54 +/- 0.7 microM for pregnenolone and 9.61 +/- 0.8 microM for NAD+. The apparent Vmax was determined as 0.82 +/- 0.02 nmol min-1 mg-1. Under the same conditions the Km values for dehydroepiandrosterone and NAD+ were 3.3 +/- 0.2 microM and 9.63 +/- 1.6 microM, respectively, and the apparent Vmax was 0.62 +/- 0.01 nmol min-1 mg-1.
Mendoza-Hernández
G
G
Departamento de Bioquímica, Facultad de Medicina, Universidad Nacional Autónoma de México, Mexico City.
López-Solache
I
I
Rendón
J L
JL
D015096:3-Hydroxysteroid Dehydrogenases / Q000096:biosynthesis*; D000735:Androstenedione / Q000096:biosynthesis; D000818:Animals; D002467:Cell Nucleus / Q000378:metabolism; D003600:Cytosol / Q000378:metabolism; D003687:Dehydroepiandrosterone / Q000378:metabolism; D004285:Dogs; D006863:Hydrogen-Ion Concentration; D066298:In Vitro Techniques; D008297:Male; D008861:Microsomes / Q000378:metabolism; D008928:Mitochondria / Q000378:metabolism; D009243:NAD / Q000494:pharmacology; D009249:NADP / Q000494:pharmacology; D010179:Pancreas / Q000201:enzymology*; D011284:Pregnenolone / Q000378:metabolism; D011374:Progesterone / Q000096:biosynthesis
D009243:NAD; D000735:Androstenedione; D003687:Dehydroepiandrosterone; D011374:Progesterone; D009249:NADP; D011284:Pregnenolone; D015096:3-Hydroxysteroid Dehydrogenases; C031954:delta(5)-3 beta-hydroxysteroid dehydrogenase