A structure/function investigation into baboon cytochrome P450 side-chain cleavage (CYP11A1)
Date
2005-12
Authors
Journal Title
Journal ISSN
Volume Title
Publisher
Stellenbosch : University of Stellenbosch
Abstract
This study describes:
1. The cloning of baboon cytochrome P450 side-chain cleavage (CYP11A1) cDNA by in
vitro 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 and
V-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 and
Thr291 in substrate binding and enzymatic activity.
6. The proposal of a topological model of the CYP11A1 active pocket as determined by
substrate docking studies.
Description
Thesis (MSc (Biochemistry))--University of Stellenbosch, 2005.
Keywords
Dissertations -- Biochemistry, Theses -- Biochemistry, Cytochrome P-450, Mutagenesis, Adrenocortical hormone, Homology (Biology), Amino acids, Baboons -- Cytogenetics, Molecular cloning