The DOI matches its Crossref record; 71 of the 71 references it cites were found in a registry or an index.
Molecular Modeling of Human P450c17 (17α-Hydroxylase/ 17,20-Lyase): Insights into Reaction Mechanisms and Effects of Mutations
Open accessAbstract
P450c17 (17alpha-hydroxylase/17,20-lyase) catalyzes steroid 17alpha-hydroxylase and 17,20-lyase activities in the biosynthesis of androgens and estrogens. These two activities are differentially regulated in a tissue-specific and developmentally programmed manner. To visualize the active site topology of human P450c17 and to study the structural basis of its substrate specificity and catalytic selectivity, we constructed a second-generation computer-graphic model of human P450c17. The energetics of the model are comparable to those of the principal template of the model, P450BMP, as determined from its crystallographic coordinates. The protein structure analysis programs PROCHECK, WHATIF, and SurVol indicate that the predicted P450c17 structure is reasonable. The hydrophobic active site accommodates both delta4 and delta5 steroid substrates in a catalytically favorable orientation. The predicted contributions of positively charged residues to the redox-partner binding site were confirmed by site-directed mutagenesis. Molecular dynamic simulations with pregnenolone, 17-OH-pregnenolone, progesterone, and 17-OH-progesterone docked into the substrate-binding pocket demonstrated that regioselectivity of the hydroxylation reactions is determined both by proximity of hydrogens to the iron-oxo complex and by the stability of the carbon radicals generated after hydrogen abstraction. The model explains the activities of all known naturally occurring and synthetic human P450c17 mutants. The model predicted that mutation of lysine 89 would disrupt 17,20-lyase activity to a greater extent than 17alpha-hydroxylase activity; expression of a test mutant, K89N, in yeast confirmed this prediction. Hydrogen peroxide did not support catalysis of the 17,20-lyase reaction, as would be predicted by mechanisms involving a ferryl peroxide. Our present model and biochemical data suggest that both the hydroxylase and lyase activities proceed from a common steroid-binding geometry by an iron oxene mechanism. This model will facilitate studies of sex steroid synthesis and its disorders and the design of specific inhibitors useful in chemotherapy of sex steroid-dependent cancers.
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“The model identifies Arg 347 and Arg 358 and several other arginine and lysine residues in the redox partner binding site; mutations of these residues all cause varying degrees of selective loss of 17,20lyase activity (240, 279, 280, 284).”
From The Molecular Biology, Biochemistry, and Physiology of Human Steroidogenesis and Its Disorders (2011) · cites this paper as [240]
In the citing paper
“Computational modeling of P450c17 predicts the effects of all known mutations, including those with partial retention of both activities and those causing selective 17,20-lyase deficiency (240).”
From The Molecular Biology, Biochemistry, and Physiology of Human Steroidogenesis and Its Disorders (2011) · cites this paper as [240]
In the citing paper
“Basic residues in the redox-partner-binding surface of the recipient P450 are crucial for electron transfer from POR (49, 302), as shown by the mutations in human P450c17 that cause 17,20lyase deficiency (240, 280).”
From The Molecular Biology, Biochemistry, and Physiology of Human Steroidogenesis and Its Disorders (2011) · cites this paper as [240]
In the citing paper
“Genetic and biochemical studies have implicated basic residues in P450c17, including R347, R358, and perhaps R449 and K89, as important for its interaction with cytochrome b 5 (240,280,348), whereas E48 and E49 of cytochrome b 5 are required for high 17,20-lyase activity (349).”
From The Molecular Biology, Biochemistry, and Physiology of Human Steroidogenesis and Its Disorders (2011) · cites this paper as [240]
In the citing paper
“By analogy with the computationally inferred structure of P450c17 (240), Arg 356 appears to be part of the redox-partner-binding site, Val 281 appears to participate in coordinating the heme moiety, and Cys 428 is the crucial cystine residue in the hemebinding site found in all cytochrome P450 enzymes.”
From The Molecular Biology, Biochemistry, and Physiology of Human Steroidogenesis and Its Disorders (2011) · cites this paper as [240]
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Metadata
- Authors
- Auchus, R. J., & Miller, W. L.
- Affiliations
- University of California, San Francisco (US)
- Corresponding author
- Miller, Walter L.
- Venue
- Molecular Endocrinology
- Publisher
- The Endocrine Society
- Published
- 1999-07
- Volume
- 13
- Issue
- 7
- Pages
- 1169-1182
- ISSN
- 0888-8809 (ISSN-L)
- 1944-9917
- OpenAlex
- W2112100343
- Cited by
- 185
- References
- 71
- Citations by year
- 2025: 3 · 2024: 1 · 2022: 2 · 2021: 4 · 2020: 1 · 2019: 1 · 2018: 1 · 2017: 2 · 2016: 5 · 2015: 7 · 2014: 8 · 2013: 8 · 2012: 8
- FWCI
- 6.63
- Type
- journal article
- Language
- en
- Open access
- bronze
- doi.org
- Locations
- Molecular Endocrinology (publishedVersion)
- PubMed (publishedVersion)
- Funders
- National Institutes of Health (P41 RP-01081, 5r01dk042154-09)
- University of California, San Francisco
- National Institute of Diabetes and Digestive and Kidney Diseases (DK-37922, DK 42154, R01DK037922)
- Eunice Kennedy Shriver National Institute of Child Health and Human Development (U54-HD34449, U54HD034449)
- Topics
- Physical Sciences › Chemistry › Inorganic Chemistry › Metal-Catalyzed Oxygenation Mechanisms
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Protein Structure and Dynamics
- Physical Sciences › Chemistry › Organic Chemistry › Steroid Chemistry and Biochemistry
- Keywords
- CYP17, 17α-hydroxylase/17,20-lyase, steroid hydroxylation, steroidogenesis, androgen production, estrogen synthesis, active site structure, substrate specificity, regioselectivity, hydrogen abstraction, pregnenolone, progesterone, molecular docking, molecular dynamics simulation
- MeSH terms
- Amino Acid Sequence, Bacterial Proteins, Binding Sites, Cytochrome P-450 Enzyme System, Heme, Humans, Mixed Function Oxygenases, Models, Molecular, Molecular Sequence Data, Mutation, NADPH-Ferrihemoprotein Reductase, Pregnenolone, Protein Conformation, Steroid 17-alpha-Hydroxylase, Mutagenesis, Site-Directed, Sequence Homology, Amino Acid
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