17 Beta-hydroxysteroid oxidoreductase activity: age-dependent profile in rat liver and kinetic properties of the hepatic microsomal enzyme in relation to cytochrome P450-dependent steroid hydroxylation.
The functional relationship between the microsomal cytochrome P450 and 17 beta-hydroxysteroid oxidoreductase (HSOR) enzymes involved in steroid metabolism was investigated in rat liver. In male and female rat hepatic microsomes the NADPH-dependent conversion of androstenedione (AD) to testosterone (T) was approx. 4-fold greater at 6 weeks of age than in 1 week old animals. In hepatic microsomes from 15 week old rats the activity of the HSOR pathway was greater in males than in females (1.51 compared to 0.80 nmol T formed/min/mg protein). However, oestradiol administration to intact adult male rats did not decrease HSOR activity. Thus, androgen is not essential for maintenance of HSOR enzymes. Instead, it is likely that irreversible androgen imprinting of the HSOR enzyme occurs during the prepubertal period. The in vitro characteristics of HSOR activity were also assessed. The Km for NADH-dependent reduction of AD to T was 9.2 microM and the Vmax was 3.0 nmol/min/mg protein but the NAD-mediated formation of AD from T did not follow Michaelis-Menton kinetics. pH markedly influenced HSOR-mediated AD/T interconversion with 17-ketosteroid reduction facilitated at low pH, and 17 beta-hydroxysteroid dehydrogenation about 2-fold more efficient at pH 8.0 than at pH 5.5. Product steroid activation of HSOR activity was noted. 17 beta-Hydroxysteroids, including T and oestradiol, activated the rate of conversion of AD to T and 17-ketosteroids such as oestrone and AD activated the NAD-dependent dehydrogenation of T. Activation was not observed at low steroid substrate concentrations so that it was not possible to analyse this phenomenon by a conventional kinetic approach.
Murray
M
M
Department of Medicine, University of Sydney, Westmead Hospital, NSW, Australia.
Horsfield
B P
BP
D015067:17-Hydroxysteroid Dehydrogenases / Q000378:metabolism*; D000367:Age Factors; D000375:Aging / Q000502:physiology*; D000735:Androstenedione / Q000378:metabolism; D000818:Animals; D004958:Estradiol / Q000494:pharmacology; D005260:Female; D006863:Hydrogen-Ion Concentration; D006914:Hydroxysteroids / Q000378:metabolism; D007700:Kinetics; D008297:Male; D008862:Microsomes, Liver / Q000201:enzymology*; D009249:NADP / Q000494:pharmacology; D051381:Rats; D011919:Rats, Inbred Strains; D012737:Sex Factors; D013254:Steroid 17-alpha-Hydroxylase / Q000378:metabolism*; D013739:Testosterone / Q000096:biosynthesis
D006914:Hydroxysteroids; D013739:Testosterone; D000735:Androstenedione; D004958:Estradiol; D009249:NADP; D015067:17-Hydroxysteroid Dehydrogenases; D013254:Steroid 17-alpha-Hydroxylase