A structure/function investigation into baboon cytochrome P450 side-chain cleavage (CYP11A1)

[摘要] This study describes:1. The cloning of baboon cytochrome P450 side-chain cleavage (CYP11A1) cDNA by invitro site-directed mutagenesis.2. The identification and sequencing of three baboon CYP11A1 mutants: CYP11A1a,CYP11A1b and CYP11A1c.3. The expression and characterisation of baboon and human CYP11A1 cDNA,CYP11A1a, CYP11A1b and CYP11A1c in nonsteroidogenic COS-1 cells. The Km andV-values for the metabolism of 25-hydroxycholesterol were determined.4. The construction of the first homology model of CYP11A1, using both mammalian(CYP2C5) and bacterial (CYP102) cytochromes P450 crystal structures as templates.5. A structure/function study into the role of the amino acid residues Ile98, Lys103 andThr291 in substrate binding and enzymatic activity.6. The proposal of a topological model of the CYP11A1 active pocket as determined bysubstrate docking studies.

[发布机构] Stellenbosch University