Efficiency of the sulfate pathway in comparison to the Δ4- and Δ5-pathway of steroidogenesis in the porcine testis

• Published in: The Journal of steroid biochemistry and molecular biology 2018-05, Vol.179, p.64-72
• Main Authors: Klymiuk, M.C., Neunzig, J., Bernhardt, R., Sánchez-Guijo, A., Hartmann, M.F., Wudy, S.A., Schuler, G.
• Format: Article
• Language: English
• Subjects: CYP17A1; Cytochrome b5; Pig; Pregnenolone; Pregnenolone sulfate; Progesterone
• ISSN: 0960-0760; 1879-1220
• DOI: 10.1016/j.jsbmb.2017.10.017

•P5S is not a quantitatively important substrate of porcine CYP17A1.•bovine b5 stimulated 17α-hydroxylation of P4 and P5 in a porcine CYP17A1/bovine CPR-system.•experimental conditions considerably influenced P5 and P4 metabolism by CYP17A1. Sulfonated steroids are increasingly recognized as a circulating reservoir of precursors for the local production of active steroids in certain target tissues. As an alternative to sulfonation of unconjugated steroids by cytosolic sulfotransferases, their direct formation from sulfonated precursors has been described. However, productivity and physiological relevance of this sulfate pathway of steroidogenesis are still widely unclear. Applying the porcine testis as a model, conversion of pregnenolone sulfate (P5S, sulfate pathway) by CYP17A1 was assessed in comparison to the parallel conversions of pregnenolone (P5, Δ5-pathway) and progesterone (P4, Δ4-pathway). To characterize conversions in the virtual absence of competing enzyme activities, in a first series of experiments porcine recombinant CYP17A1 was incubated with the respective substrate in the presence of bovine recombinant cytochrome P450 oxidoreductase (CPR) and cytochrome b5 (b5). Moreover, porcine testicular microsomal fractions were used as a source of homologous CYP17A1, CPR and b5. Invariably 17α-hydroxylation of P5S was, if at all, only minimal and no formation of dehydroepiandrosterone sulfate from P5S was detectable. Consistent with earlier studies porcine CYP17A1 efficiently metabolized P4 and P5 in both assay systems. Metabolism of P4 and P5 by testicular microsomal protein varied substantially between the five animals tested. In conclusion, a physiologically relevant sulfate pathway for the production of C19-steroids from P5S via CYP17A1 is very unlikely in the porcine testis.