Determination of functional effects of mutations in the steroid 21-hydroxylase gene (CYP21) using recombinant vaccinia virus

Maria Teresa Tusie-Luna, Paula Traktman, Perrin C. White

Research output: Contribution to journalArticle

186 Citations (Scopus)

Abstract

Steroid 21-hydroxylase (P450c21) is absent or defective in more than 90% of patients with congenital adrenal hyperplasia. This disorder of cortisol biosynthesis occurs in a wide spectrum of clinical severity; specific mutations in the 21-hydroxylase gene (CYP21) have been found in association with particular clinical phenotypes. To determine the functional effects of mutations causing amino acid substitutions, normal P450c21 and three mutagenized P450c21 enzymes were expressed at high levels in cultured COS-1 cells using recombinant vaccinia virus. A single amino acid substitution (Val281 → Leu) present in patients with mild "nonclassical" 21-hydroxylase deficiency resulted in an enzyme with 20-50% of normal activity. A mutation (Ile172 → Asn) identified in patients with the "simple virilizing" form (poor cortisol synthesis but adequate aldosterone synthesis) resulted in an enzyme with less than 2% of normal activity. Finally, a cluster mutation (Ile-Val-Glu-Met234,238 → Asn-Glu-Glu-Lys) found in a patient with severe "salt wasting" 21-hydroxylase deficiency (inadequate aldosterone synthesis) results in an enzyme with no detectable activity. These data indicate that the severity of 21-hydroxylase deficiency correlates with the degree of enzymatic compromise.

Original languageEnglish (US)
Pages (from-to)20916-20922
Number of pages7
JournalJournal of Biological Chemistry
Volume265
Issue number34
StatePublished - Dec 5 1990

Fingerprint

Steroid 21-Hydroxylase
Vaccinia virus
Viruses
Genes
Mutation
Enzymes
Amino Acid Substitution
Aldosterone
Hydrocortisone
Substitution reactions
Congenital Adrenal Hyperplasia
COS Cells
Amino Acids
Biosynthesis
Cultured Cells
Salts
Phenotype
Association reactions
Congenital adrenal hyperplasia due to 21 hydroxylase deficiency

ASJC Scopus subject areas

  • Biochemistry

Cite this

Determination of functional effects of mutations in the steroid 21-hydroxylase gene (CYP21) using recombinant vaccinia virus. / Tusie-Luna, Maria Teresa; Traktman, Paula; White, Perrin C.

In: Journal of Biological Chemistry, Vol. 265, No. 34, 05.12.1990, p. 20916-20922.

Research output: Contribution to journalArticle

@article{0dc13f13b95d429684723b5f6f2768e4,
title = "Determination of functional effects of mutations in the steroid 21-hydroxylase gene (CYP21) using recombinant vaccinia virus",
abstract = "Steroid 21-hydroxylase (P450c21) is absent or defective in more than 90{\%} of patients with congenital adrenal hyperplasia. This disorder of cortisol biosynthesis occurs in a wide spectrum of clinical severity; specific mutations in the 21-hydroxylase gene (CYP21) have been found in association with particular clinical phenotypes. To determine the functional effects of mutations causing amino acid substitutions, normal P450c21 and three mutagenized P450c21 enzymes were expressed at high levels in cultured COS-1 cells using recombinant vaccinia virus. A single amino acid substitution (Val281 → Leu) present in patients with mild {"}nonclassical{"} 21-hydroxylase deficiency resulted in an enzyme with 20-50{\%} of normal activity. A mutation (Ile172 → Asn) identified in patients with the {"}simple virilizing{"} form (poor cortisol synthesis but adequate aldosterone synthesis) resulted in an enzyme with less than 2{\%} of normal activity. Finally, a cluster mutation (Ile-Val-Glu-Met234,238 → Asn-Glu-Glu-Lys) found in a patient with severe {"}salt wasting{"} 21-hydroxylase deficiency (inadequate aldosterone synthesis) results in an enzyme with no detectable activity. These data indicate that the severity of 21-hydroxylase deficiency correlates with the degree of enzymatic compromise.",
author = "Tusie-Luna, {Maria Teresa} and Paula Traktman and White, {Perrin C.}",
year = "1990",
month = "12",
day = "5",
language = "English (US)",
volume = "265",
pages = "20916--20922",
journal = "Journal of Biological Chemistry",
issn = "0021-9258",
publisher = "American Society for Biochemistry and Molecular Biology Inc.",
number = "34",

}

TY - JOUR

T1 - Determination of functional effects of mutations in the steroid 21-hydroxylase gene (CYP21) using recombinant vaccinia virus

AU - Tusie-Luna, Maria Teresa

AU - Traktman, Paula

AU - White, Perrin C.

PY - 1990/12/5

Y1 - 1990/12/5

N2 - Steroid 21-hydroxylase (P450c21) is absent or defective in more than 90% of patients with congenital adrenal hyperplasia. This disorder of cortisol biosynthesis occurs in a wide spectrum of clinical severity; specific mutations in the 21-hydroxylase gene (CYP21) have been found in association with particular clinical phenotypes. To determine the functional effects of mutations causing amino acid substitutions, normal P450c21 and three mutagenized P450c21 enzymes were expressed at high levels in cultured COS-1 cells using recombinant vaccinia virus. A single amino acid substitution (Val281 → Leu) present in patients with mild "nonclassical" 21-hydroxylase deficiency resulted in an enzyme with 20-50% of normal activity. A mutation (Ile172 → Asn) identified in patients with the "simple virilizing" form (poor cortisol synthesis but adequate aldosterone synthesis) resulted in an enzyme with less than 2% of normal activity. Finally, a cluster mutation (Ile-Val-Glu-Met234,238 → Asn-Glu-Glu-Lys) found in a patient with severe "salt wasting" 21-hydroxylase deficiency (inadequate aldosterone synthesis) results in an enzyme with no detectable activity. These data indicate that the severity of 21-hydroxylase deficiency correlates with the degree of enzymatic compromise.

AB - Steroid 21-hydroxylase (P450c21) is absent or defective in more than 90% of patients with congenital adrenal hyperplasia. This disorder of cortisol biosynthesis occurs in a wide spectrum of clinical severity; specific mutations in the 21-hydroxylase gene (CYP21) have been found in association with particular clinical phenotypes. To determine the functional effects of mutations causing amino acid substitutions, normal P450c21 and three mutagenized P450c21 enzymes were expressed at high levels in cultured COS-1 cells using recombinant vaccinia virus. A single amino acid substitution (Val281 → Leu) present in patients with mild "nonclassical" 21-hydroxylase deficiency resulted in an enzyme with 20-50% of normal activity. A mutation (Ile172 → Asn) identified in patients with the "simple virilizing" form (poor cortisol synthesis but adequate aldosterone synthesis) resulted in an enzyme with less than 2% of normal activity. Finally, a cluster mutation (Ile-Val-Glu-Met234,238 → Asn-Glu-Glu-Lys) found in a patient with severe "salt wasting" 21-hydroxylase deficiency (inadequate aldosterone synthesis) results in an enzyme with no detectable activity. These data indicate that the severity of 21-hydroxylase deficiency correlates with the degree of enzymatic compromise.

UR - http://www.scopus.com/inward/record.url?scp=0025696003&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0025696003&partnerID=8YFLogxK

M3 - Article

VL - 265

SP - 20916

EP - 20922

JO - Journal of Biological Chemistry

JF - Journal of Biological Chemistry

SN - 0021-9258

IS - 34

ER -