Mutations of P450c21 (steroid 21-hydroxylase) at Cys428, Val281, and Ser268 result in complete, partial, or no loss of enzymatic activity, respectively.
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Steroid 21-hydroxylase deficiency is the major cause of congenital adrenal hyperplasia (CAH), a common genetic disease. To define the relationship between gene mutations and enzyme deficiency, we generated missense mutations of the 21-hydroxylase cDNA at three different sites and characterized the mutant proteins after expressing them in cultured mammalian and yeast cells. Among them, Ser268 and Val281 have been found to be mutated in CAH patients, whereas Cys428 has been implicated as the heme ligand. Our results show mutations at these sites result in complete, partial, or no loss of the enzymatic activity. All the Cys428 mutants had neither enzymatic activity nor P450 absorption, thus supporting the notion that Cys428 is the heme ligand. All the 268-mutants exhibited the same activity as normal 21-hydroxylase, demonstrating that the clinically observed Ser268----Thr change represents a polymorphism rather than the cause of the enzyme deficiency. The 281-mutants had normal Km but greatly reduced Vmax values that also paralleled the reduction in the heme content, in the order Val281 (normal, 100%) greater than Ile281 (50%) greater than Leu281 (20%) greater than Thr281 (10%). Our findings suggest that the methyl group at the beta-carbon of Val281 is required for heme incorporation and consequently enzymatic activity.
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V281L does not alter the affinity of the enzyme for substrate but drastically reduces its Vmax.
“This mutation does not alter the affinity of the enzyme for substrate but drastically reduces its V max (444).”
From The Molecular Biology, Biochemistry, and Physiology of Human Steroidogenesis and Its Disorders (2011) · cites this paper as [444]
I172N reduces enzymatic activity to <5% of normal and is associated with the SV form of the disorder (mutation Group B).
“( I172N)(154) reduces enzymatic activity to <5% of normal and is associated with the SV form of the disorder (mutation Group B)(107,109,221).”
From Congenital Adrenal Hyperplasia—Current Insights in Pathophysiology, Diagnostics, and Management (2022) · cites this paper as [109]
The Introduction identifies Ser268, Val281, and Cys428 as the three sites studied, and the Results (p. 520) report activity for mutants at those sites. A search for I172N, activity below 5%, and the simple-virilizing (SV) form found no passage addressing that mutation or its clinical association. (LLM, conf=0.75)
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Cites
The Carbon Monoxide-binding Pigment of Liver Microsomes
Omura, T., & Sato, R. (1964) · Journal of Biological Chemistry · doi:10.1016/s0021-9258(20)82244-3 · Cited by 11,781 · see citing works
HIGH-RESOLUTION CRYSTAL STRUCTURE OF CYTOCHROME P450-CAM
Poulos, T. L., Finzel, B. C., & Howard, A. J. (1987) · Journal of Molecular Biology · doi:10.1016/0022-2836(87)90190-2 · Cited by 1,414 · see citing works
Nebert, D. W., Nelson, D. R., Adesnik, M., Coon, M. J., Estabrook, R. W., Gonzalez, F. J., Guengerich, F. P., GUNSALUS, I. C., Johnson, E. F., KEMPER, B. W., et al. (1989) · DNA · doi:10.1089/dna.1.1989.8.1 · Cited by 606 · see citing works
Structure of human steroid 21-hydroxylase genes.
White, P. C., New, M. I., & Dupont, B. (1986) · Proceedings of the National Academy of Sciences · doi:10.1073/pnas.83.14.5111 · Cited by 569 · see citing works
Higashi, Y., Yoshioka, H., Yamane, M., Gotoh, O., & Fujii-Kuriyama, Y. (1986) · Proceedings of the National Academy of Sciences · doi:10.1073/pnas.83.9.2841 · Cited by 555 · see citing works
PROTEINASE MUTANTS OF SACCHAROMYCES CEREVISIAE
Jones, E. W. (1977) · Genetics · doi:10.1093/genetics/85.1.23 · Cited by 459 · see citing works
On the membrane topology of vertebrate cytochrome P-450 proteins.
Nelson, D. R., & Strobel, H. W. (1988) · Journal of Biological Chemistry · doi:10.1016/s0021-9258(18)68746-0 · Cited by 362 · see citing works
White, P. C., New, M. I., & Dupont, B. (1984) · Proceedings of the National Academy of Sciences · doi:10.1073/pnas.81.23.7505 · Cited by 292 · see citing works
HYDROXYLATION OF STEROIDS AT CARBON 21
Ryan, K. J., & Engel, L. L. (1957) · Journal of Biological Chemistry · doi:10.1016/s0021-9258(18)64913-0 · Cited by 276 · see citing works
Shen, A. L., Porter, T. D., Wilson, T. E., & Kasper, C. B. (1989) · Journal of Biological Chemistry · doi:10.1016/s0021-9258(18)83274-4 · Cited by 264 · see citing works
Tusie-Luna, M. T., Traktman, P., & White, P. C. (1990) · Journal of Biological Chemistry · doi:10.1016/s0021-9258(17)45304-x · Cited by 254 · see citing works
Mutation in the CYP21B gene (Ile-172-Asn) causes steroid 21-hydroxylase deficiency
Amor, M., Parker, K. L., Globerman, H., New, M. I., & White, P. C. (1988) · Proceedings of the National Academy of Sciences · doi:10.1073/pnas.85.5.1600 · Cited by 251 · see citing works
Higashi, Y., Tanae, A., Inoue, H., Hiromasa, T., & Fujii-Kuriyama, Y. (1988) · Proceedings of the National Academy of Sciences · doi:10.1073/pnas.85.20.7486 · Cited by 244 · see citing works
Characterization of frequent deletions causing steroid 21-hydroxylase deficiency
White, P. C., Vitek, A., Dupont, B., & New, M. I. (1988) · Proceedings of the National Academy of Sciences · doi:10.1073/pnas.85.12.4436 · Cited by 229 · see citing works
Deletion of complement C4 and steroid 21‐hydroxylase genes in the HLA class III region.
Carroll, M. C., Pálsdóttir, Á., Belt, K., & Porter, R. R. (1985) · The EMBO Journal · doi:10.1002/j.1460-2075.1985.tb03969.x · Cited by 209 · see citing works
Speiser, P. W., New, M. I., & White, P. C. (1988) · New England Journal of Medicine · doi:10.1056/nejm198807073190104 · Cited by 205 · see citing works
Rodrigues, N. R., Dunham, I., Yu, C. Y., Carroll, M. C., Porter, R. R., & Campbell, R. D. (1987) · The EMBO Journal · doi:10.1002/j.1460-2075.1987.tb02414.x · Cited by 182 · see citing works
Bitter, G. A., & Egan, K. M. (1984) · Gene · doi:10.1016/0378-1119(84)90002-7 · Cited by 179 · see citing works
Molecular and clinical advances in congenital adrenal hyperplasia
Miller, W. L., & Levine, L. S. (1987) · The Journal of Pediatrics · doi:10.1016/s0022-3476(87)80334-7 · Cited by 177 · see citing works
Higashi, Y., Tanae, A., Inoue, H., & Fujii-Kuriyama, Y. (1988) · Europe PMC (PubMed Central) · Cited by 173 · see citing works
A missense mutation at Ile172----Asn or Arg356----Trp causes steroid 21-hydroxylase deficiency.
Chiou, S. H., Hu, M. C., & Chung, B. C. (1990) · Journal of Biological Chemistry · doi:10.1016/s0021-9258(19)39804-7 · Cited by 149 · see citing works
Self-citationMorel, Y., André, J., Uring-Lambert, B., Hauptmann, G., Bétuel, H., Tossi, M., Forest, M. G., David, M., Bertrand, J., & Miller, W. L. (1989) · Journal of Clinical Investigation · doi:10.1172/jci113914 · Cited by 134 · see citing works
Nonsense mutation causing steroid 21-hydroxylase deficiency
Globerman, H., Amor, M., Parker, K. L., New, M. I., & White, P. C. (1988) · Journal of Clinical Investigation · doi:10.1172/jci113562 · Cited by 130 · see citing works
THE MOLECULAR GENETICS OF 21-HYDROXYLASE DEFICIENCY
Miller, W. L., & Morel, Y. (1989) · Annual Review of Genetics · doi:10.1146/annurev.ge.23.120189.002103 · Cited by 127 · see citing works
Site-directed mutageneses of rat liver cytochrome P-450d: axial ligand and heme incorporation
Shimizu, T., Hirano, K., Takahashi, T., et al. (1988) · Biochemistry · doi:10.1021/bi00411a035 · Cited by 83 · see citing works
Identification and location of .alpha.-helices in mammalian cytochromes P450
Edwards, R. J., Murray, B. P., Boobis, A. R., & Davies, D. S. (1989) · Biochemistry · doi:10.1021/bi00435a021 · Cited by 83 · see citing works
Hu, M.-C., & Chung, B.-C. (1990) · Molecular Endocrinology · doi:10.1210/mend-4-6-893 · Cited by 43 · see citing works
Self-citationWU, D.-A., Hu, M., & Chung, B. (1991) · DNA and Cell Biology · doi:10.1089/dna.1991.10.201 · Cited by 27 · see citing works
Self-citationThe Molecular Genetics Of 21-Hydroxylase Deficiency
Miller, W. L. (1989) · Annual Review of Genetics · doi:10.1146/annurev.genet.23.1.371 · Cited by 18 · see citing works
Exon 7<i>Nco</i>l Restriction Site within CYP21B (Steroid 21-Hydroxylase) Is a Normal Polymorphism
Donohoue, P. A., Neto, R. S., Collins, M. M., & Migeon, C. J. (1990) · Molecular Endocrinology · doi:10.1210/mend-4-9-1354 · Cited by 17 · see citing works
Molecular and Clinical Advances in Congenital Adrenal Hyperplasia
Miller, W., & Levine, L. S. (1987) · The Journal of Urology · doi:10.1016/s0022-5347(17)43712-8 · Cited by 5 · see citing works
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Research works printed but not linked: 1
- [18]18. White, P. C., M. I. New, and B. Dupont. 1986. Structure ofhuman steroid 21-hydroxylase genes. Proc. Natl. Acad. Sci. USA. 83:5111-5115.
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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
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
Wedell, A., Ritzén, E. M., Haglund-Stengler, B., & Luthman, H. (1992) · Proceedings of the National Academy of Sciences · doi:10.1073/pnas.89.15.7232 · Cited by 150 · see citing works
Deneux, C., Tardy, V., Dib, A., Mornet, E., Billaud, L., Charron, D., Morel, Y., & Kuttenn, F. (2001) · The Journal of Clinical Endocrinology & Metabolism · doi:10.1210/jc.86.1.207 · Cited by 144 · see citing works
Mutations in steroid 21-hydroxylase (CYP21)
White, P. C., Tusie-Luna, M.-T., New, M. I., & Speiser, P. W. (1994) · Human Mutation · doi:10.1002/humu.1380030408 · Cited by 129 · see citing works
Regioselective hydroxylation of steroid hormones by human cytochromes P450
Niwa, T., Murayama, N., Imagawa, Y., & Yamazaki, H. (2015) · Drug Metabolism Reviews · doi:10.3109/03602532.2015.1011658 · Cited by 121 · see citing works
Baumgartner-Parzer, S., Witsch-Baumgartner, M., & Hoeppner, W. (2020) · European Journal of Human Genetics · doi:10.1038/s41431-020-0653-5 · Cited by 110 · see citing works
The Complexities in Genotyping of Congenital Adrenal Hyperplasia: 21-Hydroxylase Deficiency
Pignatelli, D., Carvalho, B. L., Palmeiro, A., Barros, A., Guerreiro, S. G., & Macut, D. (2019) · Frontiers in Endocrinology · doi:10.3389/fendo.2019.00432 · Cited by 95 · see citing works
Steroid 21-hydroxylase (P450c21): a new allele and spread of mutations through the pseudogene
Wedell, A., & Luthman, H. (1993) · Human Genetics · doi:10.1007/bf00218263 · Cited by 81 · see citing works
l'Allemand, D. (2000) · Journal of Clinical Endocrinology & Metabolism · doi:10.1210/jc.85.12.4562 · Cited by 90 · see citing works
Baumgartner-Parzer, S. M., Schulze, E., Waldhäusl, W., Pauschenwein, S., Rondot, S., Nowotny, P., Meyer, K., Frisch, H., Waldhauser, F., & Vierhapper, H. (2001) · The Journal of Clinical Endocrinology & Metabolism · doi:10.1210/jcem.86.10.7898 · Cited by 74 · see citing works
Barbat, B., Bogyo, A., Raux-Demay, M.-C., Kuttenn, F., Boué, J., Simon-Bouy, B., Serre, J.-L., Boué, A., & Mornet, E. (1995) · Human Mutation · doi:10.1002/humu.1380050205 · Cited by 68 · see citing works
The N-terminal hydrophobic domain of P450c21 is required for membrane insertion and enzyme stability
Hsu, L. C., Hu, M. C., Cheng, H. C., Lu, J. C., & Chung, B. C. (1993) · Journal of Biological Chemistry · doi:10.1016/s0021-9258(18)82387-0 · Cited by 32 · see citing works
Self-citationPang, S., Carbunaru, G., Haider, A., Copeland, K. C., Chang, Y. T., Lutfallah, C., & Mason, J. I. (2003) · Clinical Endocrinology · doi:10.1046/j.1365-2265.2003.01716.x · Cited by 24 · see citing works
- Authors
- Wu, D. A., & Chung, B. C.
- Affiliations
- Institute of Molecular Biology, Academia Sinica (TW)
- ORCID
- Chung, B C
- Venue
- Journal of Clinical Investigation
- Published
- 1991-08-01
- Volume
- 88
- Issue
- 2
- Pages
- 519-523
- ISSN
- 0021-9738 (ISSN-L)
- OpenAlex
- W2163059891
- Cited by
- 80
- References
- 31
- Citations by year
- 2026: 1 · 2024: 1 · 2021: 2 · 2020: 2 · 2019: 2 · 2017: 2 · 2016: 3 · 2015: 1
- FWCI
- 1.98
- Type
- journal article
- Language
- en
- Open access
- bronze
- doi.org
- Locations
- Journal of Clinical Investigation (publishedVersion)
- PubMed (publishedVersion)
- Funders
- National Science Council
- Academia Sinica
- Topics
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Sexual Differentiation and Disorders
- Health Sciences › Medicine › Endocrinology, Diabetes and Metabolism › Adrenal Hormones and Disorders
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Clinical Biochemistry › Metabolism and Genetic Disorders
- Keywords
- 21-hydroxylase, CYP21, congenital adrenal hyperplasia, missense mutations, heme ligands, heme incorporation, enzyme activity, genetic polymorphism
- MeSH terms
- Adrenal Hyperplasia, Congenital, Amino Acid Sequence, Animals, Base Sequence, Cell Line, Heme, Humans, Kinetics, Molecular Sequence Data, Mutation, Oligonucleotides, Steroid 21-Hydroxylase, Structure-Activity Relationship, Transfection, Mutagenesis, Site-Directed, Sequence Alignment
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