Minor activities and transition state properties of the human steroid hydroxylases cytochromes P450c17 and P450c21, from reactions observed with deuterium-labeled substrates

Yoshimoto, Francis K.; Zhou, Yishan; Peng, Hwei-Ming; Stidd, David; Yoshimoto, Jennifer A.; Sharma, Kamalesh K.; Matthew, Susan; Auchus, Richard J.

doi:10.1021/bi300895w

Cited by 38Metal-Catalyzed Oxygenation MechanismsChemical Reaction MechanismsSteroid Chemistry and Biochemistry

Minor activities and transition state properties of the human steroid hydroxylases cytochromes P450c17 and P450c21, from reactions observed with deuterium-labeled substrates

Open access

Yoshimoto, Francis K., Zhou, Yishan, Peng, Hwei-Ming, Stidd, David, Yoshimoto, Jennifer A., Sharma, Kamalesh K., Matthew, Susan, Auchus, Richard J.·2012·Biochemistry

DOI 10.1021/bi300895wPMC PMC3471366

Open this paper in CoChat — free account

Abstract

The steroid hydroxylases CYP17A1 (P450c17, 17-hydroxylase/17,20-lyase) and CYP21A2 (P450c21, 21-hydroxylase) catalyze progesterone hydroxylation at one or more sites within a 2 Å radius. We probed their hydrogen atom abstraction mechanisms and regiochemical plasticity with deuterium-labeled substrates: 17-[(2)H]-pregnenolone; 17-[(2)H]-, 16α-[(2)H]-, 21,21,21-[(2)H(3)]-, and 21-[(2)H]-progesterone; and 21,21,21-[(2)H(3)]-17-hydroxyprogesterone. Product distribution and formation rates with recombinant human P450-oxidoreductase and wild-type human CYP17A1 or mutation A105L (reduced progesterone 16α-hydroxylation) and wild-type human CYP21A2 or mutation V359A (substantial progesterone 16α-hydroxylation) were used to calculate intramolecular and intermolecular kinetic isotope effects (KIEs). The intramolecular KIEs for CYP17A1 and mutation A105L were 4.1 and 3.8, respectively, at H-17 and 2.9 and 5.1, respectively, at H-16α. Mutation A105L 21-hydroxylates progesterone (5% of products), and wild-type CYP17A1 also catalyzes a trace of 21-hydroxylation, which increases with 16α-[(2)H]- and 17-[(2)H]-progesterone. The intramolecular KIEs with CYP21A2 mutation V359A and progesterone were 6.2 and 3.8 at H-21 and H-16α, respectively. Wild-type CYP21A2 also forms a trace of 16α-hydroxyprogesterone, which increased with 21,21,21-[(2)H(3)]-progesterone substrate. Competitive intermolecular KIEs paralleled the intramolecular KIE values, with (D)V values of 1.4-5.1 and (D)V/K values of 1.8-5.1 for these reactions. CYP17A1 and CYP21A2 mutation V359A both 16α-hydroxylate 16α-[(2)H]-progesterone with 33-44% deuterium retention, indicating stereochemical inversion. We conclude that human CYP17A1 has progesterone 21-hydroxylase activity and human CYP21A2 has progesterone 16α-hydroxylase activity, both of which are enhanced with deuterated substrates. The transition states for C-H bond cleavage in these hydroxylation reactions are either significantly nonlinear and/or asymmetric, and C-H bond breakage is partially rate-limiting for all reactions.

The claims and statistics of this paper haven’t been checked yet.

Run the check — free, no account needed. It reads the open-access text and checks the sources it cites.

Integrity signals

Registry

Crossref · 10.1021/bi300895w

Cites

47 of 47 references verified0 citation errors · 0 not checkable

Statistics

Claims the paper makes

The claims and statistics of this paper haven’t been checked yet.

Run the check — free, no account needed. It reads the open-access text and checks the sources it cites.

Claims other papers make about this one

Each is checked against this paper’s own text: does it say what the citing paper says it does?

21-hydroxylation of progesterone by hepatic cytochrome P450 enzymes other than CYP21A2 may permit some mineralocorticoid (11-deoxycorticosterone) synthesis.
“Moreover, 21-hydroxylation of progesterone by hepatic cytochrome P450 enzymes other than CYP21A2 may permit some mineralocorticoid (11deoxycorticosterone) synthesis (102).”
From Congenital Adrenal Hyperplasia—Current Insights in Pathophysiology, Diagnostics, and Management (2022) · cites this paper as [102]
The Results (page unavailable) report that CYP17A1, an enzyme other than CYP21A2, 21-hydroxylates progesterone to form 11-deoxycorticosterone (DOC). The Discussion (page unavailable) suggests this trace activity might matter in CYP21A2 deficiency. The paper does not establish that the enzyme is hepatic or that the reaction permits mineralocorticoid synthesis in the liver. (LLM, conf=0.87)
Experimental — accuracy not yet measured

Self-check

Percentages, sums and reported differences re-computed from the paper’s own numbers.

The self-check needs the paper’s full text.

Abiraterone and Increased Survival in Metastatic Prostate Cancer
de Bono, J. S., Logothetis, C. J., Molina, A., Fizazi, K., North, S., Chu, L., Chi, K. N., Jones, R. J., Goodman, O. B., Saad, F., et al. (2011) · New England Journal of Medicine · doi:10.1056/nejmoa1014618 · Cited by 4,407 · see citing works
The Molecular Biology, Biochemistry, and Physiology of Human Steroidogenesis and Its Disorders
Miller, W. L., & Auchus, R. J. (2011) · Endocrine Reviews · doi:10.1210/er.2010-0013 · Cited by 2,458 · see citing works
Self-citation
Congenital adrenal hyperplasia due to 21-hydroxylase deficiency*
White, P. C., & Speiser, P. W. (2000) · Endocrine Reviews · doi:10.1210/edrv.21.3.0398 · Cited by 1,432 · see citing works
Cytochrome b 5 Augments the 17,20-Lyase Activity of Human P450c17 without Direct Electron Transfer
Auchus, R. J., Lee, T. C., & Miller, W. L. (1998) · Journal of Biological Chemistry · doi:10.1074/jbc.273.6.3158 · Cited by 541 · see citing works
Self-citation
Aliphatic hydroxylation by highly purified liver microsomal cytochrome P-450. Evidence for a carbon radical intermediate
Groves, J. T., McClusky, G. A., White, R. E., & Coon, M. J. (1978) · Biochemical and Biophysical Research Communications · doi:10.1016/0006-291x(78)91643-1 · Cited by 535 · see citing works
Temperature-Dependent Isotope Effects in Soybean Lipoxygenase-1: Correlating Hydrogen Tunneling with Protein Dynamics
Knapp, M. J., Rickert, K. W., & Klinman, J. P. (2002) · Journal of the American Chemical Society · doi:10.1021/ja012205t · Cited by 509 · see citing works
Photochemical Action Spectrum of the Terminal Oxidase of Mixed Function Oxidase Systems
Cooper, D. Y., Levin, S. S., Narasimhulu, S., Rosenthal, O., & Estabrook, R. W. (1965) · Science · doi:10.1126/science.147.3656.400 · Cited by 501 · see citing works
Predicting phenotype in steroid 21-hydroxylase deficiency? Comprehensive genotyping in 155 unrelated, well defined patients from southern Germany
Krone, N., Braun, A., Roscher, A. A., Knorr, D., & Schwarz, H. P. (2000) · The Journal of Clinical Endocrinology & Metabolism · doi:10.1210/jcem.85.3.6441 · Cited by 397 · see citing works
Results of screening 1.9 million Texas newborns for 21-hydroxylase-deficient congenital adrenal hyperplasia
Therrell, B. L., Berenbaum, S. A., Manter-Kapanke, V., Simmank, J., Korman, K., Prentice, L., Gonzalez, J., & Gunn, S. (1998) · Pediatrics · doi:10.1542/peds.101.4.583 · Cited by 344 · see citing works
Structures of cytochrome P450 17A1 with prostate cancer drugs abiraterone and TOK-001
DeVore, N. M., & Scott, E. E. (2012) · Nature · doi:10.1038/nature10743 · Cited by 344 · see citing works
Ultrahigh-Quality NOE Spectra
Stonehouse, J., Adell, P., Keeler, J., & Shaka, A. J. (1994) · Journal of the American Chemical Society · doi:10.1021/ja00092a092 · Cited by 269 · see citing works
Mechanisms of cytochrome P450 substrate oxidation: MiniReview
Guengerich, F. P. (2007) · Journal of Biochemical and Molecular Toxicology · doi:10.1002/jbt.20174 · Cited by 249 · see citing works
Two Prevalent CYP17 Mutations and Genotype-Phenotype Correlations in 24 Brazilian Patients with 17-Hydroxylase Deficiency
Costa-Santos, M., Kater, C. E., & Auchus, R. J. (2004) · The Journal of Clinical Endocrinology & Metabolism · doi:10.1210/jc.2003-031021 · Cited by 224 · see citing works
Self-citation
The role of cytochrome b5 in the biosynthesis of androgens by human P450c17
Katagiri, M., Kagawa, N., & Waterman, M. R. (1995) · Archives of Biochemistry and Biophysics · doi:10.1006/abbi.1995.1173 · Cited by 223 · see citing works
Autooxidation and hydroxylation reactions of oxygenated cytochrome P-450cam.
Lipscomb, J. D., Sligar, S. G., Namtvedt, M. J., & GUNSALUS, I. C. (1976) · Journal of Biological Chemistry · doi:10.1016/s0021-9258(17)33808-5 · Cited by 216 · see citing works
Stereochemistry and deuterium isotope effects in camphor hydroxylation by the cytochrome P450cam monooxygenase system
Gelb, M. H., Heimbrook, D. C., Mälkönen, P. J., & Sligar, S. G. (1982) · Biochemistry · doi:10.1021/bi00531a026 · Cited by 192 · see citing works
Molecular Modeling of Human P450c17 (17α-Hydroxylase/ 17,20-Lyase): Insights into Reaction Mechanisms and Effects of Mutations
Auchus, R. J., & Miller, W. L. (1999) · Molecular Endocrinology · doi:10.1210/mend.13.7.0326 · Cited by 185 · see citing works
Self-citation
THE USE OF DEUTERIUM ISOTOPE EFFECTS TO PROBE THE ACTIVE SITE PROPERTIES, MECHANISM OF CYTOCHROME P450-CATALYZED REACTIONS, AND MECHANISMS OF METABOLICALLY DEPENDENT TOXICITY
Nelson, S. D., & Trager, W. F. (2003) · Drug Metabolism and Disposition · doi:10.1124/dmd.31.12.1481 · Cited by 164 · see citing works
Modulation of the activity of human 17 α-hydroxylase-17,20-lyase (CYP17) by cytochrome b5: endocrinological and mechanistic implications
Lee-Robichaud, P., Wright, J. N., Akhtar, M. E., & Akhtar, M. (1995) · Biochemical Journal · doi:10.1042/bj3080901 · Cited by 145 · see citing works
CYP17 mutation E305G causes isolated 17,20 lyase deficiency by selectively altering substrate binding
Sherbet, D. P., Tiosano, D., Kwist, K. M., Hochberg, Z., & Auchus, R. J. (2003) · Journal of Biological Chemistry · doi:10.1074/jbc.m307586200 · Cited by 131 · see citing works
Self-citation
Cytochrome b5 stimulates purified testicular microsomal cytochrome P-450 (C21 side-chain cleavage)
Onoda, M., & Hall, P. F. (1982) · Biochemical and Biophysical Research Communications · doi:10.1016/0006-291x(82)90850-6 · Cited by 126 · see citing works
Hydroxylation by the Hydroperoxy-Iron Species in Cytochrome P450 Enzymes
Chandrasena, R. E. P., Vatsis, K. P., Coon, M. J., Hollenberg, P. F., & Newcomb, M. (2003) · Journal of the American Chemical Society · doi:10.1021/ja038237t · Cited by 116 · see citing works
Isolated 17,20-lyase deficiency due to the cytochrome b5 mutation W27X
Kok, R. C., Timmerman, M. A., Wolffenbuttel, K. P., Drop, S. L. S., & de Jong, F. H. (2010) · The Journal of Clinical Endocrinology & Metabolism · doi:10.1210/jc.2008-1745 · Cited by 111 · see citing works
Progesterone 16 alpha-hydroxylase activity is catalyzed by human cytochrome P450 17 alpha-hydroxylase
Swart, P., Swart, A. C., Waterman, M. R., Estabrook, R. W., & Mason, J. I. (1993) · The Journal of Clinical Endocrinology & Metabolism · doi:10.1210/jcem.77.1.8325965 · Cited by 106 · see citing works
A missense mutation in the human cytochrome b5 gene causes 46,XY disorder of sex development due to true isolated 17,20 lyase deficiency
Idkowiak, J., Randell, T., Dhir, V., Patel, P., Shackleton, C. H. L., Taylor, N. F., Krone, N., & Arlt, W. (2012) · The Journal of Clinical Endocrinology & Metabolism · doi:10.1210/jc.2011-2413 · Cited by 104 · see citing works
Cytochrome P450 3A4-catalyzed Testosterone 6β-Hydroxylation Stereochemistry, Kinetic Deuterium Isotope Effects, and Rate-limiting Steps
Krauser, J. A., & Guengerich, F. P. (2005) · Journal of Biological Chemistry · doi:10.1074/jbc.m501854200 · Cited by 104 · see citing works
[30] Deuterium and tritium kinetic isotope effects on initial rates
Northrop, D. B. (1982) · Methods in enzymology on CD-ROM/Methods in enzymology · doi:10.1016/s0076-6879(82)87032-8 · Cited by 103 · see citing works
Kinetic Analysis of the Three-step Steroid Aromatase Reaction of Human Cytochrome P450 19A1
Sohl, C. D., & Guengerich, F. P. (2010) · Journal of Biological Chemistry · doi:10.1074/jbc.m110.123711 · Cited by 103 · see citing works
Kinetic Analysis of Oxidation of Coumarins by Human Cytochrome P450 2A6
Yun, C., Kim, K., Calcutt, M. W., & Guengerich, F. P. (2005) · Journal of Biological Chemistry · doi:10.1074/jbc.m411019200 · Cited by 90 · see citing works
Kinetic isotope effects on cytochrome P-450-catalyzed oxidation reactions. The oxidative O-dealkylation of 7-ethoxycoumarin.
Miwa, G. T., Walsh, J. S. A., & Lu, A. (1984) · Journal of Biological Chemistry · doi:10.1016/s0021-9258(17)43248-0 · Cited by 89 · see citing works
Cytochrome P450-Catalyzed Hydroxylation of Hydrocarbons: Kinetic Deuterium Isotope Effects for the Hydroxylation of an Ultrafast Radical Clock
Atkinson, J. K., Hollenberg, P. F., Ingold, K. U., Johnson, C. C., Tadic-Biadatti, M.-H. L., Newcomb, M., & Putt, D. A. (1994) · Biochemistry · doi:10.1021/bi00201a009 · Cited by 89 · see citing works
Rate-Limiting Steps in Oxidations Catalyzed by Rabbit Cytochrome P450 1A2
Guengerich, F. P., Krauser, J. A., & Johnson, W. W. (2004) · Biochemistry · doi:10.1021/bi0491393 · Cited by 86 · see citing works
Three-dimensional Structure of Steroid 21-Hydroxylase (Cytochrome P450 21A2) with Two Substrates Reveals Locations of Disease-associated Variants
Zhao, B., Li, L., Kagawa, N., Sundaramoorthy, M., Banerjee, S., Nagy, L. D., Guengerich, F. P., & Waterman, M. R. (2012) · Journal of Biological Chemistry · doi:10.1074/jbc.m111.323501 · Cited by 80 · see citing works
A Highly Active Single‐Mutation Variant of P450BM3 (CYP102A1)
Whitehouse, C. J. C., Bell, S. G., Yang, W., Yorke, J. A., Blanford, C. F., Strong, A. J. F., Morse, E. J., Bartlam, M. G., Rao, Z., & Wong, L. (2009) · ChemBioChem · doi:10.1002/cbic.200900279 · Cited by 76 · see citing works
Human Cytochrome b5 Requires Residues E48 and E49 to Stimulate the 17,20-Lyase Activity of Cytochrome P450c17
Naffin-Olivos, J. L., & Auchus, R. J. (2006) · Biochemistry · doi:10.1021/bi051623y · Cited by 66 · see citing works
Self-citation
Oxidation of Methoxyphenethylamines by Cytochrome P450 2D6
Guengerich, F. P., Miller, G. P., Hanna, I. H., Sato, H., & Martin, M. V. (2002) · Journal of Biological Chemistry · doi:10.1074/jbc.m205146200 · Cited by 61 · see citing works
Theory for the observed isotope effects from enzymic systems that form multiple products via branched reaction pathways: cytochrome P-450
Korzekwa, K. R., Trager, W. F., & Gillette, J. R. (1989) · Biochemistry · doi:10.1021/bi00449a009 · Cited by 57 · see citing works
Human Steroid Biosynthesis for the Oncologist
Auchus, M., & Auchus, R. J. (2012) · Journal of Investigative Medicine · doi:10.2310/jim.0b013e3182408567 · Cited by 56 · see citing works
Self-citation
Cytochrome P450 7A1 Cholesterol 7α-Hydroxylation
Shinkyo, R., & Guengerich, F. P. (2010) · Journal of Biological Chemistry · doi:10.1074/jbc.m110.193409 · Cited by 46 · see citing works
A single amino acid residue, Ala 105, confers 16α-hydroxylase activity to human cytochrome P450 17α-hydroxylase/17,20 lyase
Swart, A. C., Storbeck, K., & Swart, P. (2010) · The Journal of Steroid Biochemistry and Molecular Biology · doi:10.1016/j.jsbmb.2009.12.014 · Cited by 44 · see citing works
Human Cytochrome P450c17: Single Step Purification and Phosphorylation of Serine 258 by Protein Kinase A
Wang, Y., Tee, M. K., & Miller, W. L. (2010) · Endocrinology · doi:10.1210/en.2009-1247 · Cited by 31 · see citing works
High-Yield Expression of a Catalytically Active Membrane-Bound Protein: Human P450 Oxidoreductase
Sandee, D., & Miller, W. L. (2011) · Endocrinology · doi:10.1210/en.2011-0230 · Cited by 29 · see citing works
Why human cytochrome P450c21 is a progesterone 21-hydroxylase
Mizrachi, D., Wang, Z., Sharma, K. K., Gupta, M. K., Xu, K., Dwyer, C. R., & Auchus, R. J. (2011) · Biochemistry · doi:10.1021/bi102078e · Cited by 28 · see citing works
Self-citation
Isotope Effect Studies on the Cytochrome P450 Enzymes
Korzekwa, K. R., Gillette, J. R., & Trager, W. F. (1995) · Drug Metabolism Reviews · doi:10.3109/03602539509029814 · Cited by 21 · see citing works
Enzymic aromatization of deuterium labelled testosterone and androst-4-ene-3,17-dione
Holland, H. L., & Taylor, G. J. (1981) · Canadian Journal of Chemistry · doi:10.1139/v81-406 · Cited by 16 · see citing works
The Rate-determining Step in P450 C21-catalyzing Reactions in a Membrane-reconstituted System
Kominami, S. A. J., Owaki, A., Iwanaga, T., Tagashira, H., & Yamazaki, T. (2001) · Journal of Biological Chemistry · doi:10.1074/jbc.m006043200 · Cited by 12 · see citing works
Synthesis of halogenated pregnanes, mechanistic probes of steroid hydroxylases CYP17A1 and CYP21A2
Yoshimoto, F. K., Desilets, M. C., & Auchus, R. J. (2011) · The Journal of Steroid Biochemistry and Molecular Biology · doi:10.1016/j.jsbmb.2011.09.007 · Cited by 9 · see citing works
Self-citation

Flagged first, then by citations

Sorted by citations

Congenital Adrenal Hyperplasia—Current Insights in Pathophysiology, Diagnostics, and Management
Claahsen - van der Grinten, H. L., Speiser, P. W., Ahmed, S. F., Arlt, W., Auchus, R. J., Falhammar, H., Flück, C. E., Guasti, L., Huebner, A., Kortmann, B. B. M., et al. (2022) · Endocrine Reviews · doi:10.1210/endrev/bnab016 · Cited by 556 · see citing works
Self-citation
Steroid 17-hydroxylase and 17,20-lyase deficiencies, genetic and pharmacologic
Auchus, R. J. (2016) · The Journal of Steroid Biochemistry and Molecular Biology · doi:10.1016/j.jsbmb.2016.02.002 · Cited by 233 · see citing works
Self-citation
Human Cytochrome P450 Enzymes
Guengerich, F. P. (2015) · Springer eBooks · doi:10.1007/978-3-319-12108-6_9 · Cited by 150 · see citing works
Mechanism of the Third Oxidative Step in the Conversion of Androgens to Estrogens by Cytochrome P450 19A1 Steroid Aromatase
Yoshimoto, F. K., & Guengerich, F. P. (2014) · Journal of the American Chemical Society · doi:10.1021/ja508185d · Cited by 126 · see citing works
Self-citation
Structures of human steroidogenic cytochrome P450 17A1 with substrates
Petrunak, E. M., DeVore, N. M., Porubsky, P. R., & Scott, E. E. (2014) · Journal of Biological Chemistry · doi:10.1074/jbc.m114.610998 · Cited by 119 · see citing works
The diverse chemistry of cytochrome P450 17A1 (P450c17, CYP17A1)
Yoshimoto, F. K., & Auchus, R. J. (2014) · The Journal of Steroid Biochemistry and Molecular Biology · doi:10.1016/j.jsbmb.2014.11.026 · Cited by 117 · see citing works
Self-citation
Steroidogenic cytochrome P450 17A1 structure and function
Burris‐Hiday, S. D., & Scott, E. E. (2021) · Molecular and Cellular Endocrinology · doi:10.1016/j.mce.2021.111261 · Cited by 94 · see citing works
Human Cytochrome P450 21A2, the Major Steroid 21-Hydroxylase
Pallan, P. S., Wang, C., Li, L., Yoshimoto, F. K., Auchus, R. J., Waterman, M. R., Guengerich, F. P., & Egli, M. (2015) · Journal of Biological Chemistry · doi:10.1074/jbc.m115.646307 · Cited by 88 · see citing works
Self-citation
Mechanism of 17α,20-Lyase and New Hydroxylation Reactions of Human Cytochrome P450 17A1
Yoshimoto, F. K., Gonzalez, E. R., Auchus, R. J., & Guengerich, F. P. (2016) · Journal of Biological Chemistry · doi:10.1074/jbc.m116.732966 · Cited by 70 · see citing works
Self-citation
Abiraterone acetate treatment lowers 11-oxygenated androgens
Wright, C., O’Day, P., Alyamani, M., Sharifi, N., & Auchus, R. J. (2020) · European Journal of Endocrinology · doi:10.1530/eje-19-0905 · Cited by 68 · see citing works
Self-citation
Kinetic processivity of the two-step oxidations of progesterone and pregnenolone to androgens by human cytochrome P450 17A1
Gonzalez, E. R., & Guengerich, F. P. (2017) · Journal of Biological Chemistry · doi:10.1074/jbc.m117.794917 · Cited by 52 · see citing works
The post-translational regulation of 17,20 lyase activity
Miller, W. L., & Tee, M. K. (2014) · Molecular and Cellular Endocrinology · doi:10.1016/j.mce.2014.09.010 · Cited by 51 · see citing works
Functional analysis of human cytochrome P450 21A2 variants involved in congenital adrenal hyperplasia
Wang, C., Pallan, P. S., Zhang, W., Lei, L., Yoshimoto, F. K., Waterman, M. R., Egli, M., & Guengerich, F. P. (2017) · Journal of Biological Chemistry · doi:10.1074/jbc.m117.792465 · Cited by 43 · see citing works
Self-citation
The hunt for a selective 17,20 lyase inhibitor; learning lessons from nature
Bird, I. M., & Abbott, D. H. (2016) · The Journal of Steroid Biochemistry and Molecular Biology · doi:10.1016/j.jsbmb.2016.04.021 · Cited by 43 · see citing works
Catalytically Relevant Electrostatic Interactions of Cytochrome P450c17 (CYP17A1) and Cytochrome b5
Peng, H., Liu, J., Forsberg, S. E., Tran, H. T., Anderson, S. M., & Auchus, R. J. (2014) · Journal of Biological Chemistry · doi:10.1074/jbc.m114.608919 · Cited by 39 · see citing works
Self-citation
P450 Enzymes in Steroid Processing
Auchus, R. J., & Miller, W. L. (2015) · Springer eBooks · doi:10.1007/978-3-319-12108-6_12 · Cited by 39 · see citing works
Self-citation
Predicted Benign and Synonymous Variants in CYP11A1 Cause Primary Adrenal Insufficiency Through Missplicing
Maharaj, A. V., Buonocore, F., Meimaridou, E., Ruiz‐Babot, G., Guasti, L., Peng, H., Capper, C. P., Burgos-Tirado, N., Prasad, R., Hughes, C. R., et al. (2018) · Journal of the Endocrine Society · doi:10.1210/js.2018-00130 · Cited by 39 · see citing works
Self-citation
Instability of the Human Cytochrome P450 Reductase A287P Variant Is the Major Contributor to Its Antley-Bixler Syndrome-like Phenotype
McCammon, K. M., Panda, S. P., Xia, C., Kim, J.-J. P., Moutinho, D., Kranendonk, M., Auchus, R. J., Lafer, E. M., Ghosh, D., Martasek, P., et al. (2016) · Journal of Biological Chemistry · doi:10.1074/jbc.m116.716019 · Cited by 33 · see citing works
Self-citation
Mechanism of Cytochrome P450 17A1-Catalyzed Hydroxylase and Lyase Reactions
Bonomo, S., Jørgensen, F. S., & Olsen, L. (2017) · Journal of Chemical Information and Modeling · doi:10.1021/acs.jcim.6b00759 · Cited by 35 · see citing works
The Catalytic Cycle of the Antioxidant and Cancer-Associated Human NQO1 Enzyme: Hydride Transfer, Conformational Dynamics and Functional Cooperativity
Anoz‐Carbonell, E., Timson, D. J., Pey, A. L., & Medina, M. (2020) · Antioxidants · doi:10.3390/antiox9090772 · Cited by 34 · see citing works
Inherent steroid 17α,20-lyase activity in defunct cytochrome P450 17A enzymes
Gonzalez, E. R., Johnson, K. M., Pallan, P. S., Phan, T. T. N., Zhang, W., Li, L., Wawrzak, Z., Yoshimoto, F. K., Egli, M., & Guengerich, F. P. (2017) · Journal of Biological Chemistry · doi:10.1074/jbc.ra117.000504 · Cited by 32 · see citing works
Self-citation
Structural basis of the pleiotropic and specific phenotypic consequences of missense mutations in the multifunctional NAD(P)H:quinone oxidoreductase 1 and their pharmacological rescue
Pacheco‐García, J. L., Anoz‐Carbonell, E., Vaňková, P., Kannan, A., Palomino‐Morales, R., Mesa‐Torres, N., SALIDO, E. C., Man, P., Medina, M., Naganathan, A. N., et al. (2021) · Redox Biology · doi:10.1016/j.redox.2021.102112 · Cited by 28 · see citing works
Insights into Interactions of Human Cytochrome P450 17A1: A Review
Singh, H., Kumar, R., Mazumder, A., Salahuddin, S., Mazumder, R., & Abdullah, M. M. (2022) · Current Drug Metabolism · doi:10.2174/1389200223666220401093833 · Cited by 27 · see citing works
CYP154C5 Regioselectivity in Steroid Hydroxylation Explored by Substrate Modifications and Protein Engineering**
Bracco, P., Wijma, H. J., Nicolai, B., Buitrago, J. A. R., Klünemann, T., Vila, A., Schrepfer, P., Blankenfeldt, W., Janssen, D. B., & Schallmey, A. (2020) · ChemBioChem · doi:10.1002/cbic.202000735 · Cited by 26 · see citing works
Epoxidation Activities of Human Cytochromes P450c17 and P450c21
Yoshimoto, F. K., Peng, H., Zhang, H., Anderson, S. M., & Auchus, R. J. (2014) · Biochemistry · doi:10.1021/bi5011865 · Cited by 15 · see citing works
Self-citation
Human Steroid Biosynthesis
Auchus, R. J. (2014) · Elsevier eBooks · doi:10.1016/b978-0-12-397175-3.00008-9 · Cited by 10 · see citing works
Self-citation
Rapid kinetic methods to dissect steroidogenic cytochrome P450 reaction mechanisms
Yoshimoto, F. K., & Auchus, R. J. (2015) · The Journal of Steroid Biochemistry and Molecular Biology · doi:10.1016/j.jsbmb.2015.10.005 · Cited by 9 · see citing works
Self-citation
Effects of fluorine substitution on substrate conversion by cytochromes P450 17A1 and 21A2
Vogt, C. D., Bart, A. G., Yadav, R., Scott, E. E., & Aubé, J. (2021) · Organic & Biomolecular Chemistry · doi:10.1039/d1ob01178b · Cited by 7 · see citing works
Functional characterization of the G162R and D216H genetic variants of human CYP17A1
Capper, C. P., Liu, J., McIntosh, L., Larios, J., Johnson, M. D., Hollenberg, P. F., Osawa, Y., Auchus, R. J., & Rae, J. M. (2017) · The Journal of Steroid Biochemistry and Molecular Biology · doi:10.1016/j.jsbmb.2017.12.002 · Cited by 5 · see citing works
Self-citation
Assessment of in silico antioxidant, anti-inflammatory, and antidiabetic activites of ximenia Americana L. Olacaceae
Dahiru, M. M., Alfa, M. B., Abubakar, M. A., & Abdulllahi, A. P. (2024) · Advances in Medical Pharmaceutical and Dental Research · doi:10.21622/ampdr.2024.04.1.735 · Cited by 5 · see citing works
Characterization, In Silico Antimalarial, Antiinflammatory, Antioxidant, and ADMET Assessment of Neonauclea excelsa Merr.
Musa, N., Dahiru, M. M., & Badgal, E. B. (2024) · Sciences of Pharmacy · doi:10.58920/sciphar0302232 · Cited by 4 · see citing works
Salt-Losing 21-Hydroxylase Deficiency Caused by Double Homozygosity for Two “Mild” Mutations
Ilany, J., Liu, J., Welsch, C., Reznik‐Wolf, H., Levy‐Lahad, E., & Auchus, R. J. (2020) · The Journal of Clinical Endocrinology & Metabolism · doi:10.1210/clinem/dgaa875 · Cited by 4 · see citing works
Self-citation
Simplified approaches to kinetic isotope effects in cytochrome P450-catalyzed reactions and relevance to deuterated drugs
Guengerich, F. P. (2026) · Drug Metabolism Reviews · doi:10.1080/03602532.2026.2657290 · Cited by 2 · see citing works
Similar Rates of Second Electron Transfer and Single-Turnover Dehydroepiandrosterone Formation for Oxyferrous Human Cytochrome P450 17A1 (Steroid 17-Hydroxylase/17,20-lyase)-17-hydroxypregnenolone Complex with Either Human Cytochrome P450-Oxidoreductase or Human Cytochrome b5
Im, S., Peng, H., Waskell, L. A., & Auchus, R. J. (2025) · Biochemistry · doi:10.1021/acs.biochem.5c00217 · Cited by 2 · see citing works
Self-citation
CYP154C5 Regioselectivity in Steroid Hydroxylation Explored by Substrate and Protein EngineeringPreprint
Bracco, P., Wijma, H. J., Nicolai, B., Buitrago, J. A. R., Klünemann, T., Vila, A., Schrepfer, P., Blankenfeldt, W., Janssen, D. B., & Schallmey, A. (2020) · bioRxiv (Cold Spring Harbor Laboratory) · doi:10.18154/rwth-2021-00548
The Kinetic and Chemical Mechanisms of Human Cytochrome P450 17A1
Gonzalez, E. R. (2017)
Development of a high-throughput LLE-UPLC-QqQ-MS/MS method and comprehensive profiling of urinary steroid hormones in Chinese elderly adults
Ma, X., Ren, H., Zhao, L., Guo, X., Zhang, J., Liu, J., Wang, X., Wang, M., Lu, Y., Tong, S., et al. (2025) · Analytica Chimica Acta · doi:10.1016/j.aca.2025.344994
Strategies to inhibit steroid cytochrome P450 enzymes to benefit human health: development of steroid ligands for P450s 17A1, 19A1, and 8B1 to treat cancer and obesity
Ho, T. M., & Yoshimoto, F. K. (2025) · RSC Medicinal Chemistry · doi:10.1039/d5md00954e
Self-citation

A single amino acid residue, Ala 105, confers 16α-hydroxylase activity to human cytochrome P450 17α-hydroxylase/17,20 lyase
Swart, A. C., Storbeck, K., & Swart, P. (2010) · The Journal of Steroid Biochemistry and Molecular Biology · doi:10.1016/j.jsbmb.2009.12.014 · Cited by 44 · see citing works
Epoxidation Activities of Human Cytochromes P450c17 and P450c21
Auchus, R. J. (2016)
Self-citation
Kinetic Analysis of the Processivity of Human Cytochrome P450 17A1
Gonzalez, E. R., Nagy, L. D., & Guengerich, F. P. (2015) · The FASEB Journal · doi:10.1096/fasebj.29.1_supplement.722.13
Synthesis of halogenated pregnanes, mechanistic probes of steroid hydroxylases CYP17A1 and CYP21A2
Yoshimoto, F. K., Desilets, M. C., & Auchus, R. J. (2011) · The Journal of Steroid Biochemistry and Molecular Biology · doi:10.1016/j.jsbmb.2011.09.007 · Cited by 9 · see citing works
Self-citation
Human Cytochrome CYP17A1: The Structural Basis for Compromised Lyase Activity with 17-Hydroxyprogesterone
Mak, P. J., Duggal, R., Denisov, I. G., Gregory, M. C., Sligar, S. G., & Kincaid, J. R. (2018) · Journal of the American Chemical Society · doi:10.1021/jacs.8b03901 · Cited by 59 · see citing works

Authors: Yoshimoto, F. K., Zhou, Y., Peng, H.-M., Stidd, D., Yoshimoto, J. A., Sharma, K. K., Matthew, S., & Auchus, R. J.
Affiliations: University of Michigan (US)
Corresponding author: Auchus, Richard J.
ORCID: Yoshimoto, Francis K.
Zhou, Yishan
Peng, Hwei-Ming
Yoshimoto, Jennifer A.
Auchus, Richard J.
Venue: Biochemistry
Publisher: American Chemical Society (ACS)
Published: 2012-08-27
Volume: 51
Issue: 36
Pages: 7064-7077
ISSN: 0006-2960 (ISSN-L)
1520-4995
DOI: 10.1021/bi300895w
PMC: PMC3471366
OpenAlex: W1967240005
Cited by: 38
References: 47
Citations by year: 2026: 1 · 2025: 3 · 2024: 2 · 2022: 1 · 2021: 4 · 2020: 5 · 2018: 1 · 2017: 6 · 2016: 4 · 2015: 4 · 2014: 7
FWCI: 1.92
Type: journal article
Language: en
Open access: green
www.ncbi.nlm.nih.gov
Locations: Biochemistry (publishedVersion)
PubMed (publishedVersion)
Europe PMC (PubMed Central) (acceptedVersion)
PubMed Central (acceptedVersion)
Funders: National Institute of General Medical Sciences (R01-GM08659602, R01GM086596)
National Institute of Diabetes and Digestive and Kidney Diseases (DK089503, P30DK089503)
Topics: Physical Sciences › Chemistry › Inorganic Chemistry › Metal-Catalyzed Oxygenation Mechanisms
Physical Sciences › Chemistry › Organic Chemistry › Chemical Reaction Mechanisms
Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Steroid Chemistry and Biochemistry
Keywords: CYP17A1, CYP21A2, 21-hydroxylase, progesterone hydroxylation, hydrogen abstraction, kinetic isotope effects, regioselectivity, transition state, 16α-hydroxylation, 17-hydroxyprogesterone, cytochrome P450 oxidoreductase
MeSH terms: Deuterium, Humans, Kinetics, Protein Binding, Steroid 17-alpha-Hydroxylase, Steroid 21-Hydroxylase, Steroids

About these checks

Registry, retraction, Cites and statistics results are computed, not judged, with one exception: a printed reference that no index lists is read once by a model before it counts as a citation error. Claims are extracted by a model and kept only when their quote appears verbatim in the text; each cited source is then checked, with a model’s help, for whether it says what the claim relies on. Those verdicts can be wrong — read the claim’s quote and the cited source before relying on one.

Statistics and arithmetic are recomputed, not judged.

Confidence bands are shown only for fields calibrated on our annotated test set. None is calibrated yet, so no band is shown.

What counts as a claim ›How we measure confidence ›How we check this ›

Are you an author, or do you see a problem with this page? Request a review

Minor activities and transition state properties of the human steroid hydroxylases cytochromes P450c17 and P450c21, from reactions observed with deuterium-labeled substrates

Abstract

Integrity signals

Claims the paper makes

Claims other papers make about this one

Self-check

Cites

Cited by

Related

Record

About these checks