A missense mutation of Ile172 → Asn or Arg356 → Trp causes steroid 21-hydroxylase deficiency

S. H. Chiou, M. C. Hu, Bon Chu Chung

Research output: Contribution to journalArticle

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Abstract

Congenital adrenal hyperplasia (CAH) is a common recessive genetic disease caused mainly by steroid 21-hydroxylase (P450c21) deficiency. Many forms of CAH exist resulting from various mutations of the CYP21B gene. We sequenced CYP21B cDNA from a normal person and its genes from a patient with simple virilizing CAH. When comparing several CYP21B sequences, we found it was polymorphic. In the patient, a single base substitution replaced Ile172 (ATC) with Asn (AAC) in one allele while Arg356 (CGG) was converted to Trp (TGG) in the other. A normal P450c21 cDNA clone was transfected into COS-1 cells to produce 21-hydroxylase activity toward its substrates, progesterone and 17-hydroxyprogesterone. Mutants corresponding to Asn172 or Trp356 mutation were constructed by site-directed mutagenesis of the normal c21 cDNA clone. They failed to produce active enzyme toward either substrate upon transfection into COS-1 cells, demonstrating that these mutations caused CAH. Aligning sequences with other P450s, Ile172 could be located in the membrane anchoring domain and Arg356 in the substrate-binding site of P450c21. Both mutations are present in the CYP21A1P pseudogene, suggesting that they may be transferred from CYP21A1P by gene conversion events.

Original languageEnglish
Pages (from-to)3549-3552
Number of pages4
JournalJournal of Biological Chemistry
Volume265
Issue number6
Publication statusPublished - Mar 23 1990

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Steroid 21-Hydroxylase
Congenital Adrenal Hyperplasia
Missense Mutation
Complementary DNA
Mutation
COS Cells
Substrates
Genes
17-alpha-Hydroxyprogesterone
Gene Conversion
Mutagenesis
Clone Cells
Inborn Genetic Diseases
Pseudogenes
Progesterone
Substitution reactions
Site-Directed Mutagenesis
Binding Sites
Membranes
Transfection

All Science Journal Classification (ASJC) codes

  • Biochemistry
  • Molecular Biology
  • Cell Biology

Cite this

A missense mutation of Ile172 → Asn or Arg356 → Trp causes steroid 21-hydroxylase deficiency. / Chiou, S. H.; Hu, M. C.; Chung, Bon Chu.

In: Journal of Biological Chemistry, Vol. 265, No. 6, 23.03.1990, p. 3549-3552.

Research output: Contribution to journalArticle

Chiou, S. H. ; Hu, M. C. ; Chung, Bon Chu. / A missense mutation of Ile172 → Asn or Arg356 → Trp causes steroid 21-hydroxylase deficiency. In: Journal of Biological Chemistry. 1990 ; Vol. 265, No. 6. pp. 3549-3552.
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