Production, purification, and functional analysis of recombinant human and mouse 17ß-hydroxysteroid dehydrogenase type 7
Por:
Törn S., Nokelainen P., Kurkela R., Pulkka A., Menjivar M., Ghosh S., Coca-Prados M., Peltoketo H., Isomaa V., Vihko P.
Publicada:
1 ene 2003
Resumen:
17?-Hydroxysteroid dehydrogenases (17HSDs) have a central role in the regulation of the biological activity of sex steroid hormones. There is increasing evidence that in addition to their importance in gonads, these hormones also have substantial metabolic roles in a variety of peripheral tissues. In the present study, the cDNA of human 17HSD type 7 was cloned. In silico, the gene corresponding to the cDNA was localized on chromosome 1q23, close to the locus of hereditary prostate cancer 1 (HPC1) (1q24-25) and primary open-angle glaucoma (GLC1A) (1q23-25). Further, a pseudogene was found on chromosome 1q44, close to the locus of predisposing for early-onset prostate cancer (PCaP) (1q42.2-43). Both human (h17HSD7) and mouse 17HSD type 7 (m17HSD7) were for the first time produced as recombinant proteins and purified for functional analyses. Further, kinetic parameters and specific activities were described. h17HSD7 converted estrone (E1) to a more potent estrogen, estradiol (E2), and dihydrotestosterone (DHT), a potent androgen, to an estrogenic metabolite 5?-androstane-3?, 17?-diol (3?A-diol) equally, thereby catalyzing the reduction of the keto group in either 17- or 3-position of the substrate. Minor 3?HSD-like activity towards progesterone (P) and 20-hydroxyprogesterone (20-OH-P), leading to the inactivation of P by h17HSD7, was also detected. m17HSD7 efficiently catalyzed the reaction from E1 to E2 and moderately converted DHT to an inactive metabolite 5?-androstane-3?,17?-diol (3?A-diol) and to an even lesser degree 3?A-diol. The mouse enzyme did not metabolize P or 20-OH-P. The expression of 17HSD type 7 was observed widely in human tissues, most distinctly in adrenal gland, liver, lung, and thymus. Based on the enzymatic characteristics and tissue distribution, we conclude that h17HSD7 might be an intracrine regulator of steroid metabolism, fortifying the estrogenic milieu in peripheral tissues. © 2003 Elsevier Science (USA). All rights reserved.
Filiaciones:
Törn S.:
Biocenter Oulu, Res. Ctr. for Molec. Endocrinology, University of Oulu, POB 5000, FIN-90014 Oulu, Finland
Nokelainen P.:
Biocenter Oulu, Res. Ctr. for Molec. Endocrinology, University of Oulu, POB 5000, FIN-90014 Oulu, Finland
Department of Dermatology, Stanford University, Stanford, CA 94305, United States
Kurkela R.:
Biocenter Oulu, Res. Ctr. for Molec. Endocrinology, University of Oulu, POB 5000, FIN-90014 Oulu, Finland
Pulkka A.:
Biocenter Oulu, Res. Ctr. for Molec. Endocrinology, University of Oulu, POB 5000, FIN-90014 Oulu, Finland
Menjivar M.:
Biocenter Oulu, Res. Ctr. for Molec. Endocrinology, University of Oulu, POB 5000, FIN-90014 Oulu, Finland
Department of Biology, Faculty of Chemistry, Univ. Nac. Auton. de Mex., CP 04510, Mexico City, Mexico
Ghosh S.:
Dept. of Ophthalmol./Visual Science, Yale University School of Medicine, New Haven, CT 06510, United States
Coca-Prados M.:
Dept. of Ophthalmol./Visual Science, Yale University School of Medicine, New Haven, CT 06510, United States
Peltoketo H.:
Biocenter Oulu, Res. Ctr. for Molec. Endocrinology, University of Oulu, POB 5000, FIN-90014 Oulu, Finland
Faculty of Medicine, Imperial College, IRDB, Du Cane Road, London W12 0NN, United Kingdom
Isomaa V.:
Biocenter Oulu, Res. Ctr. for Molec. Endocrinology, University of Oulu, POB 5000, FIN-90014 Oulu, Finland
Vihko P.:
Biocenter Oulu, Res. Ctr. for Molec. Endocrinology, University of Oulu, POB 5000, FIN-90014 Oulu, Finland
|