Abstract
A mutation at the surface of the substrate access channel which dramatically decreases the affinity for some fatty acids in P450(BM-3) was discovered by random mutagenesis. The mutation introduced, proline-25 to glutamine, is in close proximity to the arginine-47 residue thought to be responsible for the initial docking of fatty acid substrates. The P25Q mutant displays an affinity for palmitate which is approximately 100-fold weaker than the wild-type enzyme. In addition to its altered substrate affinity, P25Q also exhibits altered hydroxylation specificity and carbon monoxide recombination kinetics in the substrate-free form.
Original language | English (US) |
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Pages (from-to) | 213-218 |
Number of pages | 6 |
Journal | FEBS Letters |
Volume | 414 |
Issue number | 2 |
DOIs | |
State | Published - Sep 8 1997 |
Keywords
- Carbon monoxide geminate kinetics
- Random mutagenesis
- Regiospecificity
- Substrate binding
ASJC Scopus subject areas
- Biophysics
- Structural Biology
- Biochemistry
- Molecular Biology
- Genetics
- Cell Biology