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J Steroid Biochem Mol Biol. 1995 Aug;54(3-4):185-92.

Rat steroid 5 alpha-reductase kinetic characteristics: extreme pH-dependency of the type II isozyme in prostate and epididymis homogenates.

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1
Department of Medicine, University of Hospital, Nijmegen, The Netherlands.

Abstract

Reevaluating the assay for rat steroid 5 alpha-reductase isozymes in prostate and epididymis homogenates we encountered an extreme pH-dependency of the type II isozyme. The time-course of the metabolism of testosterone (T) to 17 beta-hydroxy-5 alpha-androstan-3-one (DHT) at acidic pH shows an initial burst when the homogenate is not brought to pH before the start of the incubation. Therefore, the rat type II 5 alpha-reductase isozyme does not follow Michaelian law under these conditions making a single time point measurement invalid. Assessing the pH-optimum of 5 alpha-reduction in both rat prostate and epididymis homogenates we found a strong substrate dependency: at high substrate concentrations a pH-optimum for the type II isozyme of pH 5.0 was found, whereas at lower concentrations pH 5.5 is optimal. Establishing Vmax (maximum velocities) and Km (affinity constants) for the 5 alpha-reduction of T at pH 4.5-8.0, the efficiency optimum Vmax/Km appeared to be pH 5.5 in both prostate and epididymis homogenates. Specifically at acidic pH these kinetic characteristics of the type II isozyme vary many-fold. Discrepancies in literature concerning 5 alpha-reductase characteristics can, at least in part, be attributed to the choice of optimal pH, or to pH shifts during the assay.

PMID:
7662592
DOI:
10.1016/0960-0760(95)00125-j
[Indexed for MEDLINE]

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