Skip to main content
NCBI home page
Proceedings of the National Academy of Sciences of the United States of America logoLink to Proceedings of the National Academy of Sciences of the United States of America
. 1992 Jun 1;89(11):4996–5000. doi: 10.1073/pnas.89.11.4996

Mutations in the human CYP11B2 (aldosterone synthase) gene causing corticosterone methyloxidase II deficiency.

L Pascoe 1, K M Curnow 1, L Slutsker 1, A Rösler 1, P C White 1
PMCID: PMC49215  PMID: 1594605

Abstract

Corticosterone methyloxidase II (CMO-II) deficiency is an autosomal recessive disorder of aldosterone biosynthesis, characterized by an elevated ratio of 18-hydroxycorticosterone to aldosterone in serum. It is genetically linked to the CYP11B1 and CYP11B2 genes that, respectively, encode two cytochrome P450 isozymes, P450XIB1 and P450XIB2. Whereas P450XIB1 only catalyzes hydroxylation at position 11 beta of 11-deoxycorticosterone and 11-deoxycortisol, P450XIB2 catalyzes the synthesis of aldosterone from deoxycorticosterone, a process that successively requires hydroxylation at positions 11 beta and 18 and oxidation at position 18. To determine the molecular genetic basis of CMO-II deficiency, seven kindreds of Iranian-Jewish origin were studied in which members suffered from CMO-II deficiency. No mutations were found in the CYP11B1 genes, but two candidate mutations, R181W and V386A, were found in the CYP11B2 genes. When these mutations were individually introduced into CYP11B2 cDNA and expressed in cultured cells, R181W reduced 18-hydroxylase and abolished 18-oxidase activities but left 11 beta-hydroxylase activity intact, whereas V386A caused a small but consistent reduction in the production of 18-hydroxycorticosterone. All individuals affected with CMO-II deficiency were homozygous for both mutations, whereas eight asymptomatic subjects were homozygous for R181W alone and three were homozygous for V386A alone. These findings confirm that P450XIB2 is the major enzyme mediating oxidation at position 18 in the adrenal and suggest that a small amount of residual activity undetectable in in vitro assays is sufficient to synthesize normal amounts of aldosterone.

Full text

PDF
4996

Images in this article

4998Image
on p.4998
4998Image
on p.4998
4999Image
on p.4999

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Agarwal A. K., Monder C., Eckstein B., White P. C. Cloning and expression of rat cDNA encoding corticosteroid 11 beta-dehydrogenase. J Biol Chem. 1989 Nov 15;264(32):18939–18943. [PubMed] [Google Scholar]
  2. Andersson S., Davis D. L., Dahlbäck H., Jörnvall H., Russell D. W. Cloning, structure, and expression of the mitochondrial cytochrome P-450 sterol 26-hydroxylase, a bile acid biosynthetic enzyme. J Biol Chem. 1989 May 15;264(14):8222–8229. [PubMed] [Google Scholar]
  3. Casanova J. L., Pannetier C., Jaulin C., Kourilsky P. Optimal conditions for directly sequencing double-stranded PCR products with sequenase. Nucleic Acids Res. 1990 Jul 11;18(13):4028–4028. doi: 10.1093/nar/18.13.4028. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Chomczynski P., Sacchi N. Single-step method of RNA isolation by acid guanidinium thiocyanate-phenol-chloroform extraction. Anal Biochem. 1987 Apr;162(1):156–159. doi: 10.1006/abio.1987.9999. [DOI] [PubMed] [Google Scholar]
  5. Chua S. C., Szabo P., Vitek A., Grzeschik K. H., John M., White P. C. Cloning of cDNA encoding steroid 11 beta-hydroxylase (P450c11). Proc Natl Acad Sci U S A. 1987 Oct;84(20):7193–7197. doi: 10.1073/pnas.84.20.7193. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Cohen T., Theodor R., Rösler A. Selective hypoaldosteronism in Iranian Jews: An autosomal recessive trait. Clin Genet. 1977 Jan;11(1):25–30. doi: 10.1111/j.1399-0004.1977.tb01273.x. [DOI] [PubMed] [Google Scholar]
  7. Cooper D. N., Youssoufian H. The CpG dinucleotide and human genetic disease. Hum Genet. 1988 Feb;78(2):151–155. doi: 10.1007/BF00278187. [DOI] [PubMed] [Google Scholar]
  8. Curnow K. M., Tusie-Luna M. T., Pascoe L., Natarajan R., Gu J. L., Nadler J. L., White P. C. The product of the CYP11B2 gene is required for aldosterone biosynthesis in the human adrenal cortex. Mol Endocrinol. 1991 Oct;5(10):1513–1522. doi: 10.1210/mend-5-10-1513. [DOI] [PubMed] [Google Scholar]
  9. Fuerst T. R., Niles E. G., Studier F. W., Moss B. Eukaryotic transient-expression system based on recombinant vaccinia virus that synthesizes bacteriophage T7 RNA polymerase. Proc Natl Acad Sci U S A. 1986 Nov;83(21):8122–8126. doi: 10.1073/pnas.83.21.8122. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Globerman H., Rösler A., Theodor R., New M. I., White P. C. An inherited defect in aldosterone biosynthesis caused by a mutation in or near the gene for steroid 11-hydroxylase. N Engl J Med. 1988 Nov 3;319(18):1193–1197. doi: 10.1056/NEJM198811033191804. [DOI] [PubMed] [Google Scholar]
  11. Kater C. E., Biglieri E. G., Rost C. R., Schambelan M., Hirai J., Chang B. C., Brust N. The constant plasma 18-hydroxycorticosterone to aldosterone ratio: an expression of the efficacy of corticosterone methyloxidase type II activity in disorders with variable aldosterone production. J Clin Endocrinol Metab. 1985 Feb;60(2):225–228. doi: 10.1210/jcem-60-2-225. [DOI] [PubMed] [Google Scholar]
  12. Kawainoto T., Mitsuuchi Y., Ohnishi T., Ichikawa Y., Yokoyama Y., Sumimoto H., Toda K., Miyahara K., Kuribayashi I., Nakao K. Cloning and expression of a cDNA for human cytochrome P-450aldo as related to primary aldosteronism. Biochem Biophys Res Commun. 1990 Nov 30;173(1):309–316. doi: 10.1016/s0006-291x(05)81058-7. [DOI] [PubMed] [Google Scholar]
  13. Kawamoto T., Mitsuuchi Y., Toda K., Miyahara K., Yokoyama Y., Nakao K., Hosoda K., Yamamoto Y., Imura H., Shizuta Y. Cloning of cDNA and genomic DNA for human cytochrome P-45011 beta. FEBS Lett. 1990 Sep 3;269(2):345–349. doi: 10.1016/0014-5793(90)81190-y. [DOI] [PubMed] [Google Scholar]
  14. Matsukawa N., Nonaka Y., Ying Z., Higaki J., Ogihara T., Okamoto M. Molecular cloning and expression of cDNAS encoding rat aldosterone synthase: variants of cytochrome P-450(11 beta). Biochem Biophys Res Commun. 1990 May 31;169(1):245–252. doi: 10.1016/0006-291x(90)91460-a. [DOI] [PubMed] [Google Scholar]
  15. Mornet E., Dupont J., Vitek A., White P. C. Characterization of two genes encoding human steroid 11 beta-hydroxylase (P-450(11) beta). J Biol Chem. 1989 Dec 15;264(35):20961–20967. [PubMed] [Google Scholar]
  16. Ogishima T., Mitani F., Ishimura Y. Isolation of aldosterone synthase cytochrome P-450 from zona glomerulosa mitochondria of rat adrenal cortex. J Biol Chem. 1989 Jul 5;264(19):10935–10938. [PubMed] [Google Scholar]
  17. Ogishima T., Shibata H., Shimada H., Mitani F., Suzuki H., Saruta T., Ishimura Y. Aldosterone synthase cytochrome P-450 expressed in the adrenals of patients with primary aldosteronism. J Biol Chem. 1991 Jun 15;266(17):10731–10734. [PubMed] [Google Scholar]
  18. Okamura T., John M. E., Zuber M. X., Simpson E. R., Waterman M. R. Molecular cloning and amino acid sequence of the precursor form of bovine adrenodoxin: evidence for a previously unidentified COOH-terminal peptide. Proc Natl Acad Sci U S A. 1985 Sep;82(17):5705–5709. doi: 10.1073/pnas.82.17.5705. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Picado-Leonard J., Voutilainen R., Kao L. C., Chung B. C., Strauss J. F., 3rd, Miller W. L. Human adrenodoxin: cloning of three cDNAs and cycloheximide enhancement in JEG-3 cells. J Biol Chem. 1988 Mar 5;263(7):3240–3244. [PubMed] [Google Scholar]
  20. Rauh W., Levine L. S., Gottesdiener K., New M. I. Mineralocorticoids, salt balance and blood pressure after prolonged ACTH administration in juvenile hypertension. Klin Wochenschr. 1978;56 (Suppl 1):161–167. doi: 10.1007/BF01477468. [DOI] [PubMed] [Google Scholar]
  21. Rösler A., Rabinowitz D., Theodor R., Ramirez L. C., Ulick S. The nature of the defect in a salt-wasting disorder in Jews of Iran. J Clin Endocrinol Metab. 1977 Feb;44(2):279–291. doi: 10.1210/jcem-44-2-279. [DOI] [PubMed] [Google Scholar]
  22. Rösler A. The natural history of salt-wasting disorders of adrenal and renal origin. J Clin Endocrinol Metab. 1984 Oct;59(4):689–700. doi: 10.1210/jcem-59-4-689. [DOI] [PubMed] [Google Scholar]
  23. Solish S. B., Picado-Leonard J., Morel Y., Kuhn R. W., Mohandas T. K., Hanukoglu I., Miller W. L. Human adrenodoxin reductase: two mRNAs encoded by a single gene on chromosome 17cen----q25 are expressed in steroidogenic tissues. Proc Natl Acad Sci U S A. 1988 Oct;85(19):7104–7108. doi: 10.1073/pnas.85.19.7104. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Tusie-Luna M. T., Traktman P., White P. C. Determination of functional effects of mutations in the steroid 21-hydroxylase gene (CYP21) using recombinant vaccinia virus. J Biol Chem. 1990 Dec 5;265(34):20916–20922. [PubMed] [Google Scholar]
  25. ULICK S., GAUTIER E., VETTER K. K., MARKELLO J. R., YAFFE S., LOWE C. U. AN ALDOSTERONE BIOSYNTHETIC DEFECT IN A SALT-LOSING DISORDER. J Clin Endocrinol Metab. 1964 Jul;24:669–672. doi: 10.1210/jcem-24-7-669. [DOI] [PubMed] [Google Scholar]
  26. Ulick S. Diagnosis and nomenclature of the disorders of the terminal portion of the aldosterone biosynthetic pathway. J Clin Endocrinol Metab. 1976 Jul;43(1):92–96. doi: 10.1210/jcem-43-1-92. [DOI] [PubMed] [Google Scholar]
  27. Veldhuis J. D., Kulin H. E., Santen R. J., Wilson T. E., Melby J. C. Inborn error in the terminal step of aldosterone biosynthesis. Corticosterone methyl oxidase tpe II deficiency in a North American pedigree. N Engl J Med. 1980 Jul 17;303(3):117–121. doi: 10.1056/NEJM198007173030301. [DOI] [PubMed] [Google Scholar]
  28. White P. C., Dupont J., New M. I., Leiberman E., Hochberg Z., Rösler A. A mutation in CYP11B1 (Arg-448----His) associated with steroid 11 beta-hydroxylase deficiency in Jews of Moroccan origin. J Clin Invest. 1991 May;87(5):1664–1667. doi: 10.1172/JCI115182. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Winship P. R. An improved method for directly sequencing PCR amplified material using dimethyl sulphoxide. Nucleic Acids Res. 1989 Feb 11;17(3):1266–1266. doi: 10.1093/nar/17.3.1266. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Yanagibashi K., Haniu M., Shively J. E., Shen W. H., Hall P. The synthesis of aldosterone by the adrenal cortex. Two zones (fasciculata and glomerulosa) possess one enzyme for 11 beta-, 18-hydroxylation, and aldehyde synthesis. J Biol Chem. 1986 Mar 15;261(8):3556–3562. [PubMed] [Google Scholar]

Articles from Proceedings of the National Academy of Sciences of the United States of America are provided here courtesy of National Academy of Sciences

RESOURCES