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Deconvoluting the electron transfer pathway in flavocytochrome P450 BM3

ReferenceB16808
Principal Investigator / Supervisor Professor Andrew Munro
Co-Investigators /
Co-Supervisors
Professor Nigel Scrutton
Institution University of Leicester
DepartmentChemistry
Funding typeResearch
Value (£) 200,428
StatusCompleted
TypeResearch Grant
Start date 21/01/2002
End date 20/07/2005
Duration42 months

Abstract

B. megaterium flavocytochrome P450 BM3 is a soluble, self-contained P450, where a mammalian-like cytochrome P450 reductase is fused to a fatty acid hydroxylase P450 in a single polypeptide. It is an excellent system by which mechanism and electron transfer (et) rates can be determined in a eukaryotic-like (class II) P450 system. Previous work has inadequately/incorrectly described the et processes, and an impossible pathway for P450 et was proposed. We will examine et pathway/kinetics, using sited-directed mutagenesis to determine roles of key amino acids, and stopped-flow absorption/fluorescence, T-jump and laser flash photolysis to determine rates for individual catalytic steps. Newly developed stopped-flow analysis systems will permit analysis of rates of up to 3000/s and allow us to deconvolute accurately mechanism and kinetics in this model P450 system.

Summary

unavailable
Committee Closed Committee - Biomolecular Sciences (BMS)
Research TopicsX – not assigned to a current Research Topic
Research PriorityX – Research Priority information not available
Research Initiative X - not in an Initiative
Funding SchemeX – not Funded via a specific Funding Scheme
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