Mutation of proline 409 to arginine in the meander region of cytochrome P450c17 causes severe 17α-hydroxylase deficiency

Abstract

We elucidated the molecular basis of 17α-hydroxylase deficiency in a Chinese patient with male pseudohermaphroditism. The patient is a compound heterozygote, carrying two different mutant alleles in the CYP17 gene. The first mutation, g.6333-6341delGACTCTTTCA, located in exon 8, was reported in a Thai patient living in a rural village in Thailand. We suggest that g. 6333-6341del-GACTCTTTCA may be a prevalent mutation causing P450c17 deficiency in Southeast Asia. The second mutation is a mlssense mutation, g. 5582C>G, located in exon 7, changing the codon 409 from CCG to CGG, and changing the coded amino acid from proline to arginine, i.e., P409R. This proline residue is conserved in P450c17 of other species and other human P450 proteins. Site-directed mutagenesis, in vitro expression, and functional analysis of the P409R mutant in COS-1 cells show that it has a complete lack of 17α-hydroxylase activity. The pro. line residue probably causes a turn in the meander region of P450c17, and we hypothesize, by comparison to homologous proteins, that the change in the protein conformation may abolish heine incorporation or may prevent P450c17 from interacting with electron donors. © 2001 Academic Press.