help button home button Endocrine Society Endocrinology
HOME HELP FEEDBACK SUBSCRIPTIONS ARCHIVE SEARCH TABLE OF CONTENTS
This Article
Right arrow Full Text (PDF)
Right arrow Purchase Article
Right arrow View Shopping Cart
Right arrow Alert me when this article is cited
Right arrow Alert me if a correction is posted
Right arrow Citation Map
Services
Right arrow Email this article to a friend
Right arrow Similar articles in this journal
Right arrow Similar articles in PubMed
Right arrow Alert me to new issues of the journal
Right arrow Download to citation manager
Right arrow Request Copyright Permission
Citing Articles
Right arrow Citing Articles via HighWire
Right arrow Citing Articles via Google Scholar
Google Scholar
Right arrow Articles by Lin, D.
Right arrow Articles by Miller, W. L.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Lin, D.
Right arrow Articles by Miller, W. L.

Endocrinology, Vol 132, 2498-2506, Copyright © 1993 by Endocrine Society

ARTICLES

Steroid 17 alpha-hydroxylase and 17,20-lyase activities of P450c17: contributions of serine106 and P450 reductase

D Lin, SM Black, Y Nagahama and WL Miller
Department of Pediatrics, University of California, San Francisco 94143- 0978.

Cytochrome P450c17 (EC 1.14.99.9) catalyzes both 17 alpha-hydroxylase and 17,20-lyase activities in mammalian steroidogenesis and also has some 16 alpha-hydroxylase activity. The ratio of 17 alpha-hydroxylase to 17,20-lyase activity differs in the adrenal and testis and is developmentally regulated at adrenarche, but the nature of the enzyme's active site and the differential regulation of its two principal activities are unknown. The spontaneous human P450c17 mutation Ser106-- >Pro eliminates all enzymatic activity. We used site-directed mutagenesis to construct expression vectors for the conservative P450c17 mutations Ser106-->Thr and Ser106-->Ala. When expressed in transfected COS-1 cells, these mutants retain only 20-30% of the 17 alpha-hydroxylase and 17,20-lyase activities, but retain 60% of the 16 alpha-hydroxylase activity of the Ser106 wild type. Thus, the amino acid occupying position 106 greatly affects enzymatic activity. Ser is found at position 106 in P450c17 in all mammals and birds studied, but the corresponding residue (position 112) in fish (trout) is Thr. Both the trout Thr112 wild type and a Thr112-->Ser trout mutant had equivalent 16 alpha-hydroxylase, 17 alpha-hydroxylase, and 17,20-lyase activities, although these were only 5%, 5%, and 10%, respectively, of human Ser106. To catalyze its activities, P450c17 must receive electrons from NADPH via a flavoprotein termed P450 reductase. We examined the influence of the ratio of P450c17 to P450 reductase on enzymatic activity by cotransfecting COS-1 cells with varying amounts of vectors expressing each protein. The endogenous P450 reductase of COS-1 cells was sufficient to confer maximal 17 alpha-hydroxylase activity. P450 reductase produced from the transfected expression vector did not increase the conversion of [14C]progesterone to 17 alpha- or 16 alpha-hydroxyprogesterone, indicating that the endogenous immunodetectable P450 reductase of COS-1 cells was sufficient to confer maximal 17 alpha-hydroxylase activity. By contrast, the additional P450 reductase produced by the expression vector increased 17,20-lyase activity about 3-fold. Thus, the availability of reducing equivalents is a crucial factor in regulating 17,20-lyase activity. P450 reductase also increased the 17,20-lyase activity of the Thr106 and Ala106 mutants. These data suggest that the essential role of Ser106 is in the active site, rather than in interacting with P450 reductase, and that electron transfer may play an important role in regulating the 17,20- lyase activity of P450c17.


This article has been cited by other articles:


Home page
 EndocrinologyHome page
M. K. Tee, Q. Dong, and W. L. Miller
Pathways Leading to Phosphorylation of P450c17 and to the Posttranslational Regulation of Androgen Biosynthesis
Endocrinology, May 1, 2008; 149(5): 2667 - 2677.
[Abstract] [Full Text] [PDF]

Home page
 J EndocrinolHome page
E. Samandari, P. Kempna, J.-M. Nuoffer, G. Hofer, P. E Mullis, and C. E Fluck
Human adrenal corticocarcinoma NCI-H295R cells produce more androgens than NCI-H295A cells and differ in 3{beta}-hydroxysteroid dehydrogenase type 2 and 17,20 lyase activities
J. Endocrinol., December 1, 2007; 195(3): 459 - 472.
[Abstract] [Full Text] [PDF]

Home page
 J. Neurosci.Home page
J.-L. Do-Rego, S. Acharjee, J. Y. Seong, L. Galas, D. Alexandre, P. Bizet, A. Burlet, H. B. Kwon, V. Luu-The, G. Pelletier, et al.
Vasotocin and mesotocin stimulate the biosynthesis of neurosteroids in the frog brain.
J. Neurosci., June 21, 2006; 26(25): 6749 - 6760.
[Abstract] [Full Text] [PDF]

Home page
 Mol. Endocrinol.Home page
N. Huang, A. Dardis, and W. L. Miller
Regulation of Cytochrome b5 Gene Transcription by Sp3, GATA-6, and Steroidogenic Factor 1 in Human Adrenal NCI-H295A Cells
Mol. Endocrinol., August 1, 2005; 19(8): 2020 - 2034.
[Abstract] [Full Text] [PDF]

Home page
 Biol. Reprod.Home page
Y. Jo, S. R. King, S. A. Khan, and D. M. Stocco
Involvement of Protein Kinase C and Cyclic Adenosine 3',5'-Monophosphate-Dependent Kinase in Steroidogenic Acute Regulatory Protein Expression and Steroid Biosynthesis in Leydig Cells
Biol Reprod, August 1, 2005; 73(2): 244 - 255.
[Abstract] [Full Text] [PDF]

Home page
 J. Clin. Endocrinol. Metab.Home page
S. Dharia, A. Slane, M. Jian, M. Conner, A. J. Conley, R. M. Brissie, and C. R. Parker Jr.
Effects of Aging on Cytochrome B5 Expression in the Human Adrenal Gland
J. Clin. Endocrinol. Metab., July 1, 2005; 90(7): 4357 - 4361.
[Abstract] [Full Text] [PDF]

Home page
 EndocrinologyHome page
W. L. Miller
Minireview: Regulation of Steroidogenesis by Electron Transfer
Endocrinology, June 1, 2005; 146(6): 2544 - 2550.
[Abstract] [Full Text] [PDF]

Home page
 J. Biol. Chem.Home page
A. V. Pandey and W. L. Miller
Regulation of 17,20 Lyase Activity by Cytochrome b5 and by Serine Phosphorylation of P450c17
J. Biol. Chem., April 8, 2005; 280(14): 13265 - 13271.
[Abstract] [Full Text] [PDF]

Home page
 J. Biol. Chem.Home page
W.-H. Yang and S. R Hammes
Xenopus laevis CYP17 Regulates Androgen Biosynthesis Independent of the Cofactor Cytochrome b5
J. Biol. Chem., March 18, 2005; 280(11): 10196 - 10201.
[Abstract] [Full Text] [PDF]

Home page
 CirculationHome page
S. Nakamura, M. Yoshimura, M. Nakayama, T. Ito, Y. Mizuno, E. Harada, T. Sakamoto, Y. Saito, K. Nakao, H. Yasue, et al.
Possible Association of Heart Failure Status With Synthetic Balance Between Aldosterone and Dehydroepiandrosterone in Human Heart
Circulation, September 28, 2004; 110(13): 1787 - 1793.
[Abstract] [Full Text] [PDF]

Home page
 Mol. Cell. Biol.Home page
S. R. Bair and S. H. Mellon
Deletion of the Mouse P450c17 Gene Causes Early Embryonic Lethality
Mol. Cell. Biol., June 15, 2004; 24(12): 5383 - 5390.
[Abstract] [Full Text] [PDF]

Home page
 Mol. Endocrinol.Home page
C. E. Fluck and W. L. Miller
GATA-4 and GATA-6 Modulate Tissue-Specific Transcription of the Human Gene for P450c17 by Direct Interaction with Sp1
Mol. Endocrinol., May 1, 2004; 18(5): 1144 - 1157.
[Abstract] [Full Text] [PDF]

Home page
 J. Clin. Endocrinol. Metab.Home page
M. Costa-Santos, C. E. Kater, E. P. Dias, and R. J. Auchus
Two Intronic Mutations Cause 17-Hydroxylase Deficiency by Disrupting Splice Acceptor Sites: Direct Demonstration of Aberrant Splicing and Absent Enzyme Activity by Expression of the Entire CYP17 Gene in HEK-293 Cells
J. Clin. Endocrinol. Metab., January 1, 2004; 89(1): 43 - 48.
[Abstract] [Full Text] [PDF]

Home page
 J. Clin. Endocrinol. Metab.Home page
M. Costa-Santos, C. E. Kater, and R. J. Auchus
Two Prevalent CYP17 Mutations and Genotype-Phenotype Correlations in 24 Brazilian Patients with 17-Hydroxylase Deficiency
J. Clin. Endocrinol. Metab., January 1, 2004; 89(1): 49 - 60.
[Abstract] [Full Text] [PDF]

Home page
 J. Biol. Chem.Home page
D. P. Sherbet, D. Tiosano, K. M. Kwist, Z. Hochberg, and R. J. Auchus
CYP17 Mutation E305G Causes Isolated 17,20-Lyase Deficiency by Selectively Altering Substrate Binding
J. Biol. Chem., December 5, 2003; 278(49): 48563 - 48569.
[Abstract] [Full Text] [PDF]

Home page
 Reproductive SciencesHome page
K. S. Rehman, B. R. Carr, and W. E. Rainey
Profiling the Steroidogenic Pathway in Human Fetal and Adult Adrenals
Reproductive Sciences, September 1, 2003; 10(6): 372 - 380.
[Abstract] [PDF]

Home page
 J. Clin. Endocrinol. Metab.Home page
C. E. Fluck, W. L. Miller, and R. J. Auchus
The 17, 20-Lyase Activity of Cytochrome P450c17 from Human Fetal Testis Favors the {Delta}5 Steroidogenic Pathway
J. Clin. Endocrinol. Metab., August 1, 2003; 88(8): 3762 - 3766.
[Abstract] [Full Text] [PDF]

Home page
 J. Clin. Endocrinol. Metab.Home page
M. O. Muench, J. V. Ratcliffe, M. Nakanishi, H. Ishimoto, and R. B. Jaffe
Isolation of Definitive Zone and Chromaffin Cells Based upon Expression of CD56 (Neural Cell Adhesion Molecule) in the Human Fetal Adrenal Gland
J. Clin. Endocrinol. Metab., August 1, 2003; 88(8): 3921 - 3930.
[Abstract] [Full Text] [PDF]

Home page
 J. Clin. Endocrinol. Metab.Home page
M. Quinkler, C. Bumke-Vogt, B. Meyer, V. Bahr, W. Oelkers, and S. Diederich
The Human Kidney Is a Progesterone-Metabolizing and Androgen-Producing Organ
J. Clin. Endocrinol. Metab., June 1, 2003; 88(6): 2803 - 2809.
[Abstract] [Full Text] [PDF]

Home page
 J. Biol. Chem.Home page
W.-H. Yang, L. B. Lutz, and S. R. Hammes
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
J. Biol. Chem., March 7, 2003; 278(11): 9552 - 9559.
[Abstract] [Full Text] [PDF]

Home page
 J. Clin. Endocrinol. Metab.Home page
G. Guercio, M. A. Rivarola, E. Chaler, M. Maceiras, and A. Belgorosky
Relationship between the Growth Hormone/Insulin-Like Growth Factor-I Axis, Insulin Sensitivity, and Adrenal Androgens in Normal Prepubertal and Pubertal Girls
J. Clin. Endocrinol. Metab., March 1, 2003; 88(3): 1389 - 1393.
[Abstract] [Full Text] [PDF]

Home page
 J. Biol. Chem.Home page
A. V. Pandey, S. H. Mellon, and W. L. Miller
Protein Phosphatase 2A and Phosphoprotein SET Regulate Androgen Production by P450c17
J. Biol. Chem., January 24, 2003; 278(5): 2837 - 2844.
[Abstract] [Full Text] [PDF]

Home page
 EndocrinologyHome page
W. Arlt, J. W. M. Martens, M. Song, J. T. Wang, R. J. Auchus, and W. L. Miller
Molecular Evolution of Adrenarche: Structural and Functional Analysis of P450c17 from Four Primate Species
Endocrinology, December 1, 2002; 143(12): 4665 - 4672.
[Abstract] [Full Text] [PDF]

Home page
 J. Clin. Endocrinol. Metab.Home page
G. Guercio, M. A. Rivarola, E. Chaler, M. Maceiras, and A. Belgorosky
Relationship between the GH/IGF-I Axis, Insulin Sensitivity, and Adrenal Androgens in Normal Prepubertal and Pubertal Boys
J. Clin. Endocrinol. Metab., March 1, 2002; 87(3): 1162 - 1169.
[Abstract] [Full Text] [PDF]

Home page
 Proc. Natl. Acad. Sci. USAHome page
L. B. Lutz, L. M. Cole, M. K. Gupta, K. W. Kwist, R. J. Auchus, and S. R. Hammes
Evidence that androgens are the primary steroids produced by Xenopus laevis ovaries and may signal through the classical androgen receptor to promote oocyte maturation
PNAS, November 9, 2001; (2001) 241471598.
[Abstract] [Full Text] [PDF]

Home page
 J. Clin. Endocrinol. Metab.Home page
M. K. Gupta, D. H. Geller, and R. J. Auchus
Pitfalls in Characterizing P450c17 Mutations Associated with Isolated 17,20-Lyase Deficiency
J. Clin. Endocrinol. Metab., September 1, 2001; 86(9): 4416 - 4423.
[Abstract] [Full Text] [PDF]

Home page
 Mol. Endocrinol.Home page
C. J. Lin, J. W. M. Martens, and W. L. Miller
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
Mol. Endocrinol., August 1, 2001; 15(8): 1277 - 1293.
[Abstract] [Full Text] [PDF]

Home page
 J. Clin. Endocrinol. Metab.Home page
R. Azziz, L. M. Fox, H. A. Zacur, C. R. Parker Jr., and L. R. Boots
Adrenocortical Secretion of Dehydroepiandrosterone in Healthy Women: Highly Variable Response to Adrenocorticotropin
J. Clin. Endocrinol. Metab., June 1, 2001; 86(6): 2513 - 2517.
[Abstract] [Full Text] [PDF]

Home page
 EndocrinologyHome page
M.-C. Huang and W. L. Miller
Creation and Activity of COS-1 Cells Stably Expressing the F2 Fusion of the Human Cholesterol Side-Chain Cleavage Enzyme System
Endocrinology, June 1, 2001; 142(6): 2569 - 2576.
[Abstract] [Full Text] [PDF]

Home page
 Endocr. Rev.Home page
L. Ibáñez, J. DiMartino-Nardi, N. Potau, and P. Saenger
Premature Adrenarche--Normal Variant or Forerunner of Adult Disease?
Endocr. Rev., December 1, 2000; 21(6): 671 - 696.
[Abstract] [Full Text]

Home page
 J. Clin. Endocrinol. Metab.Home page
J. W. M. Martens, D. H. Geller, W. Arlt, R. J. Auchus, V. S. Ossovskaya, H. Rodriguez, A. Dunaif, and W. L. Miller
Enzymatic Activities of P450c17 Stably Expressed in Fibroblasts from Patients with the Polycystic Ovary Syndrome
J. Clin. Endocrinol. Metab., November 1, 2000; 85(11): 4338 - 4346.
[Abstract] [Full Text]

Home page
 Mol. Endocrinol.Home page
N. A. Compagnone, P. Zhang, J.-L. Vigne, and S. H. Mellon
Novel Role for the Nuclear Phosphoprotein SET in Transcriptional Activation of P450c17 and Initiation of Neurosteroidogenesis
Mol. Endocrinol., June 1, 2000; 14(6): 875 - 888.
[Abstract] [Full Text]

Home page
 Mol. Endocrinol.Home page
R. J. Auchus and W. L. Miller
Molecular Modeling of Human P450c17 (17{alpha}-Hydroxylase/ 17,20-Lyase): Insights into Reaction Mechanisms and Effects of Mutations
Mol. Endocrinol., July 1, 1999; 13(7): 1169 - 1182.
[Abstract] [Full Text]

Home page
 J. Clin. Endocrinol. Metab.Home page
T. C. Lee, W. L. Miller, and R. J. Auchus
Medroxyprogesterone Acetate and Dexamethasone Are Competitive Inhibitors of Different Human Steroidogenic Enzymes
J. Clin. Endocrinol. Metab., June 1, 1999; 84(6): 2104 - 2110.
[Abstract] [Full Text]

Home page
 Mol. Endocrinol.Home page
D. H. Geller, R. J. Auchus, and W. L. Miller
P450c17 Mutations R347H and R358Q Selectively Disrupt 17,20-Lyase Activity by Disrupting Interactions with P450 Oxidoreductase and Cytochrome b5
Mol. Endocrinol., January 1, 1999; 13(1): 167 - 175.
[Abstract] [Full Text]

Home page
 J. Biol. Chem.Home page
R. J. Auchus, T. C. Lee, and W. L. Miller
Cytochrome b5 Augments the 17,20-Lyase Activity of Human P450c17 without Direct Electron Transfer
J. Biol. Chem., February 6, 1998; 273(6): 3158 - 3165.
[Abstract] [Full Text] [PDF]

Home page
 J. Clin. Endocrinol. Metab.Home page
A. Biason-Lauber, E. Leiberman, and M. Zachmann
A Single Amino Acid Substitution in the Putative Redox Partner-Binding Site of P450c17 as Cause of Isolated 17,20-Lyase Deficiency
J. Clin. Endocrinol. Metab., November 1, 1997; 82(11): 3807 - 3812.
[Abstract] [Full Text] [PDF]

Home page
 J. Biol. Chem.Home page
G. C. Ibeanu, B. I. Ghanayem, P. Linko, L. Li, L. G. Pedersen, and J. A. Goldstein
Identification of Residues 99, 220, and 221 of Human Cytochrome P450 2C19 as Key Determinants of Omeprazole Hydroxylase Activity
J. Biol. Chem., May 24, 1996; 271(21): 12496 - 12501.
[Abstract] [Full Text] [PDF]

Home page
 J. Biol. Chem.Home page
W. Arlt, R. J. Auchus, and W. L. Miller
Thiazolidinediones but Not Metformin Directly Inhibit the Steroidogenic Enzymes P450c17 and 3beta -Hydroxysteroid Dehydrogenase
J. Biol. Chem., May 11, 2001; 276(20): 16767 - 16771.
[Abstract] [Full Text] [PDF]

Home page
 Proc. Natl. Acad. Sci. USAHome page
L. B. Lutz, L. M. Cole, M. K. Gupta, K. W. Kwist, R. J. Auchus, and S. R. Hammes
Evidence that androgens are the primary steroids produced by Xenopus laevis ovaries and may signal through the classical androgen receptor to promote oocyte maturation
PNAS, November 20, 2001; 98(24): 13728 - 13733.
[Abstract] [Full Text] [PDF]


HOME HELP FEEDBACK SUBSCRIPTIONS ARCHIVE SEARCH TABLE OF CONTENTS
Endocrinology Endocrine Reviews J. Clin. End. & Metab.
Molecular Endocrinology Recent Prog. Horm. Res. All Endocrine Journals
Copyright © 1993 by The Endocrine Society