Note to users. If you're seeing this message, it means that your browser cannot find this page's style/presentation instructions -- or possibly that you are using a browser that does not support current Web standards. Find out more about why this message is appearing, and what you can do to make your experience of our site the best it can be.

Science 5 December 1986:
Vol. 234. no. 4781, pp. 1258 - 1261
DOI: 10.1126/science.3535074
Prev | Table of Contents | Next

Articles

Science, Vol 234, Issue 4781, 1258-1261
Copyright © 1986 by American Association for the Advancement of Science

articles

Expression of bovine 17 alpha-hydroxylase cytochrome P-450 cDNA in nonsteroidogenic (COS 1) cells

MX Zuber, ER Simpson, and MR Waterman

Cortisol production requires the activity of only 17 alpha-hydroxylase, whereas the formation of sex steroids requires both 17 alpha-hydroxylase and 17,20-lyase activities. Studies in reconstituted enzyme systems have suggested that a single steroid hydroxylase, 17 alpha-hydroxylase cytochrome P-450 (P-450(17) alpha), catalyzes both activities. By expression of bovine adrenocortical P-450(17 alpha) in COS 1 (transformed monkey kidney) cells, which normally contain no detectable P-450(17) alpha, it has now been established in situ that a single polypeptide chain does catalyze both the 17 alpha-hydroxylase and the 17,20-lyase reactions. This heterologous system supports 17 alpha-hydroxylation of pregnenolone and progesterone with equal efficiency, but catalyzes about five times as much 17,20-lyase activity when 17 alpha-hydroxypregnenolone is the substrate than when 17 alpha-hydroxyprogesterone is the substrate. For these activities to be observed in COS 1 cells, newly synthesized apocytochrome P-450(17) alpha must bind heme and insert into the endoplasmic reticulum such that endogenous cytochrome P-450 reductase can support hydroxylation. Thus, COS 1 cells are a useful system for expression and study of various forms of cytochrome P-450.


THIS ARTICLE HAS BEEN CITED BY OTHER ARTICLES:
Pathways Leading to Phosphorylation of P450c17 and to the Posttranslational Regulation of Androgen Biosynthesis.
M. K. Tee, Q. Dong, and W. L. Miller (2008)
Endocrinology 149, 2667-2677
   Abstract »    Full Text »    PDF »
Phenotype-Genotype Correlation in Eight Chinese 17{alpha}-Hydroxylase/17,20 Lyase-Deficiency Patients with Five Novel Mutations of CYP17A1 Gene.
J. Yang, B. Cui, S. Sun, T. Shi, S. Zheng, Y. Bi, J. Liu, Y. Zhao, J. Chen, G. Ning, et al. (2006)
J. Clin. Endocrinol. Metab. 91, 3619-3625
   Abstract »    Full Text »    PDF »
Reciprocal expression of 17{alpha}-hydroxylase-C17,20-lyase and aromatase cytochrome P450 during bovine trophoblast differentiation: a two-cell system drives placental oestrogen synthesis..
G Schuler, G R Ozalp, B Hoffmann, N Harada, P Browne, and A J Conley (2006)
Reproduction 131, 669-679
   Abstract »    Full Text »    PDF »
17{alpha}-Hydroxylase/17,20-Lyase Deficiency Caused by a Novel Homozygous Mutation (Y27Stop) in the Cytochrome CYP17 Gene.
K. Mussig, S. Kaltenbach, F. Machicao, C. Maser-Gluth, M. F. Hartmann, S. A. Wudy, G. Schnauder, H.-U. Haring, F. J. Seif, and B. Gallwitz (2005)
J. Clin. Endocrinol. Metab. 90, 4362-4365
   Abstract »    Full Text »    PDF »
Minireview: Regulation of Steroidogenesis by Electron Transfer.
W. L. Miller (2005)
Endocrinology 146, 2544-2550
   Abstract »    Full Text »    PDF »
Regulation of 17,20 Lyase Activity by Cytochrome b5 and by Serine Phosphorylation of P450c17.
A. V. Pandey and W. L. Miller (2005)
J. Biol. Chem. 280, 13265-13271
   Abstract »    Full Text »    PDF »
Xenopus laevis CYP17 Regulates Androgen Biosynthesis Independent of the Cofactor Cytochrome b5.
W.-H. Yang and S. R Hammes (2005)
J. Biol. Chem. 280, 10196-10201
   Abstract »    Full Text »    PDF »
Overview of Steroidogenic Enzymes in the Pathway from Cholesterol to Active Steroid Hormones.
A. H. Payne and D. B. Hales (2004)
Endocr. Rev. 25, 947-970
   Abstract »    Full Text »    PDF »
Phospholipids modify substrate binding and enzyme activity of human cytochrome P450 27A1.
D. A. Murtazina, U. Andersson, I.-S. Hahn, I. Bjorkhem, G. A. S. Ansari, and I. A. Pikuleva (2004)
J. Lipid Res. 45, 2345-2353
   Abstract »    Full Text »    PDF »
Colocalization of P450c17 and Cytochrome b5 in Androgen-Synthesizing Tissues of the Human.
S. Dharia, A. Slane, M. Jian, M. Conner, A. J. Conley, and C. R. Parker Jr (2004)
Biol Reprod 71, 83-88
   Abstract »    Full Text »    PDF »
Deletion of the Mouse P450c17 Gene Causes Early Embryonic Lethality.
S. R. Bair and S. H. Mellon (2004)
Mol. Cell. Biol. 24, 5383-5390
   Abstract »    Full Text »    PDF »
Insulin Drives Transcriptional Activity of the CYP17 Gene in Primary Culturesof Swine Theca Cells.
G. Zhang and J. D. Veldhuis (2004)
Biol Reprod 70, 1600-1605
   Abstract »    Full Text »    PDF »
SUBSTRATE SPECIFICITY AND KINETIC PROPERTIES OF SEVEN HETEROLOGOUSLY EXPRESSED DOG CYTOCHROMES P450.
M. Shou, R. Norcross, G. Sandig, P. Lu, Y. Li, Y. Lin, Q. Mei, A. D. Rodrigues, and T. H. Rushmore (2003)
Drug Metab. Dispos. 31, 1161-1169
   Abstract »    Full Text »    PDF »
The 17, 20-Lyase Activity of Cytochrome P450c17 from Human Fetal Testis Favors the {Delta}5 Steroidogenic Pathway.
C. E. Fluck, W. L. Miller, and R. J. Auchus (2003)
J. Clin. Endocrinol. Metab. 88, 3762-3766
   Abstract »    Full Text »    PDF »
Xenopus laevis Ovarian CYP17 Is a Highly Potent Enzyme Expressed Exclusively in Oocytes. EVIDENCE THAT OOCYTES PLAY A CRITICAL ROLE IN XENOPUS OVARIAN ANDROGEN PRODUCTION.
W.-H. Yang, L. B. Lutz, and S. R. Hammes (2003)
J. Biol. Chem. 278, 9552-9559
   Abstract »    Full Text »    PDF »
Protein Phosphatase 2A and Phosphoprotein SET Regulate Androgen Production by P450c17.
A. V. Pandey, S. H. Mellon, and W. L. Miller (2003)
J. Biol. Chem. 278, 2837-2844
   Abstract »    Full Text »    PDF »
Combined 17{alpha}-Hydroxylase/17,20-Lyase Deficiency Caused by Phe93Cys Mutation in the CYP17 Gene.
A. Di Cerbo, A. Biason-Lauber, M. Savino, M. R. Piemontese, A. Di Giorgio, M. Perona, and A. Savoia (2002)
J. Clin. Endocrinol. Metab. 87, 898-905
   Abstract »    Full Text »    PDF »
NF-1C, Sp1, and Sp3 Are Essential for Transcription of the Human Gene for P450c17 (Steroid 17{alpha}-hydroxylase/17,20 lyase) in Human Adrenal NCI-H295A Cells.
C. J. Lin, J. W. M. Martens, and W. L. Miller (2001)
Mol. Endocrinol. 15, 1277-1293
   Abstract »    Full Text »    PDF »
In Vitro Characterization of the Oxidative Cleavage of the Octyl Side Chain of Olanexidine, a Novel Antimicrobial Agent, in Dog Liver Microsomes.
K. Umehara, S. Kudo, Y. Hirao, S. Morita, T. Ohtani, M. Uchida, and G. Miyamoto (2001)
Drug Metab. Dispos. 28, 1417-1424
   Abstract »    Full Text »
Establishment and Characterization of a Steroidogenic Human Granulosa-Like Tumor Cell Line, KGN, That Expresses Functional Follicle-Stimulating Hormone Receptor.
Y. Nishi, T. Yanase, Y.-M. Mu, K. Oba, I. Ichino, M. Saito, M. Nomura, C. Mukasa, T. Okabe, K. Goto, et al. (2001)
Endocrinology 142, 437-445
   Abstract »    Full Text »    PDF »
Enzymatic Activities of P450c17 Stably Expressed in Fibroblasts from Patients with the Polycystic Ovary Syndrome.
J. W. M. Martens, D. H. Geller, W. Arlt, R. J. Auchus, V. S. Ossovskaya, H. Rodriguez, A. Dunaif, and W. L. Miller (2000)
J. Clin. Endocrinol. Metab. 85, 4338-4346
   Abstract »    Full Text »
Interactive Stimulation by Luteinizing Hormone and Insulin of the Steroidogenic Acute Regulatory (StAR) Protein and 17{alpha}-Hydroxylase/17, 20-Lyase (CYP17) Genes in Porcine Theca Cells.
G. Zhang, J. C. Garmey, and J. D. Veldhuis (2000)
Endocrinology 141, 2735-2742
   Abstract »    Full Text »    PDF »
Characterization of the UDP-Glucuronosyltransferase 1A Locus in Lagomorphs: Evidence for Duplication of the UGT1A6 Gene.
Q. Li, G. Lamb, and R. H. Tukey (2000)
Mol. Pharmacol. 58, 89-97
   Abstract »    Full Text »
The Relationship between a Polymorphism in CYP17 with Plasma Hormone Levels and Breast Cancer.
C. A. Haiman, S. E. Hankinson, D. Spiegelman, G. A. Colditz, W. C. Willett, F. E. Speizer, K. T. Kelsey, and D. J. Hunter (1999)
Cancer Res. 59, 1015-1020
   Abstract »    Full Text »    PDF »
The tomato DWARF enzyme catalyses C-6 oxidation in brassinosteroid biosynthesis.
G. J. Bishop, T. Nomura, T. Yokota, K. Harrison, T. Noguchi, S. Fujioka, S. Takatsuto, J. D. G. Jones, and Y. Kamiya (1999)
PNAS 96, 1761-1766
   Abstract »    Full Text »    PDF »
Dehydroepiandrosterone: A potential signalling molecule for neocortical organization during development.
N. A. Compagnone and S. H. Mellon (1998)
PNAS 95, 4678-4683
   Abstract »    Full Text »    PDF »
Baculovirus-Directed Expression of Rabbit UDP-Glucuronosyltransferases in Spodoptera frugiperda Cells.
N. Nguyen and R. H. Tukey (1997)
Drug Metab. Dispos. 25, 745-749
   Abstract »    Full Text »
A Putative Binding Site for Sp1 Is Involved in Transcriptional Regulation of CYP17 Gene Expression in Bovine Ovary.
R. Borroni, Z. Liu, E. R. Simpson, and M. M. Hinshelwood (1997)
Endocrinology 138, 2011-2020
   Abstract »    Full Text »    PDF »
Male Pseudohermaphroditism Resulting From 17{alpha}-Monooxygenase (P-450C17) Deficiency in Two Unrelated Guamanians.
K. L. Jones, G. R. Freidenberg, R. Buchta, and A. Derenoncourt (1992)
Arch Pediatr Adolesc Med 146, 592-595
   Abstract »    PDF »
The neuronal growth-associated protein GAP-43 induces filopodia in non-neuronal cells.
M. Zuber, D. Goodman, L. Karns, and M. Fishman (1989)
Science 244, 1193-1195
   Abstract »    PDF »
Thiazolidinediones but Not Metformin Directly Inhibit the Steroidogenic Enzymes P450c17 and 3beta -Hydroxysteroid Dehydrogenase.
W. Arlt, R. J. Auchus, and W. L. Miller (2001)
J. Biol. Chem. 276, 16767-16771
   Abstract »    Full Text »    PDF »
Evidence that androgens are the primary steroids produced by Xenopus laevis ovaries and may signal through the classical androgen receptor to promote oocyte maturation.
L. B. Lutz, L. M. Cole, M. K. Gupta, K. W. Kwist, R. J. Auchus, and S. R. Hammes (2001)
PNAS 98, 13728-13733
   Abstract »    Full Text »    PDF »


To Advertise     Find Products

Science. ISSN 0036-8075 (print), 1095-9203 (online)