Testosterone dehydrogenase activity in koala liver: characterisation of cofactor and steroid substrate differences.
We have studied the hepatic microsomal 17beta-hydroxysteroid dehydrogenase (17beta-HSD) capacity of koala (Phascolarctos cinereus) and tammar wallaby (Macropus eugenii). A detailed comparison of the activity in hepatic fractions from koala and rat was made. Hepatic microsomal NADP-supported 17beta-HSD activity was significantly higher in koala (11.64+/-3.35 nmoles/mg protein/min), (mean+/-S.D.) than in tammar wallaby liver (1.52+/-0.79 nmoles/mg protein/min). However, when NAD was utilised as cofactor the activity was similar in both marsupial species (2.83+/-2.03 nmoles/mg protein/min, koala; 0.70+/-0.71 nmoles/mg protein/min, tammar wallaby). Data for rat indicated a cofactor preference for NAD rather than NADP (17.94+/-6.40 nmoles/mg protein/min, NAD; 2.18+/-1.04 nmoles/mg protein/min, NADP). Michaelis-Menten parameters for the kinetics of 17beta-HSD testosterone oxidation by NADP and NAD were determined in the koala. The Km for testosterone was of the order of 10.0-24.0 microM (n = 6) irrespective of the cofactor used, whilst the Km for NADP was 0.28-0.43 microM (n = 2) and for NAD was 13.9-18.5 microM (n = 2). 17beta-estradiol was found to be an inhibitor of both NAD- and NADP- supported 17beta-HSD activity. These findings indicate that NADP-mediated, but not NAD-mediated testosterone dehydrogenation is a major pathway of steroid biotransformation in koala liver; the reaction is less extensive in fractions from wallaby, human and rat. Such species-related differences in cofactor preference may contribute along with species differences in gene expression to observed rates of 17beta-HSD activity in mammals.
Stupans
I
I
Centre for Pharmaceutical Research, School of Pharmacy and Medical Sciences, University of South Australia, North Terrace, Adelaide, SA, 5000, Australia. ieva.stupans@unisa.edu.au
Kong
S
S
Kirlich
A
A
McKinnon
R A
RA
Murray
M
M
D015067:17-Hydroxysteroid Dehydrogenases / Q000378:metabolism*; D000818:Animals; D006801:Humans; D007700:Kinetics; D008394:Marsupialia / Q000502:physiology*; D008862:Microsomes, Liver / Q000201:enzymology; D009243:NAD / Q000378:metabolism; D009249:NADP / Q000378:metabolism; D010084:Oxidation-Reduction; D051381:Rats; D013379:Substrate Specificity; D013739:Testosterone / Q000378:metabolism
D009243:NAD; D013739:Testosterone; D009249:NADP; D015067:17-Hydroxysteroid Dehydrogenases