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

190 Scopus citations

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 - 1990

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology
  • Cell Biology

Fingerprint Dive into the research topics of 'Determination of functional effects of mutations in the steroid 21-hydroxylase gene (CYP21) using recombinant vaccinia virus'. Together they form a unique fingerprint.

  • Cite this