Please use this identifier to cite or link to this item:
Scopus Web of Science® Altmetric
Type: Journal article
Title: Chain length-dependent cooperativity in fatty acid binding and oxidation by cytochrome P450BM3 (CYP102A1)
Author: Rowlatt, B.
Yorke, J.
Strong, A.
Whitehouse, C.
Bell, S.
Wong, L.
Citation: Protein & Cell, 2011; 2(8):656-671
Publisher: Gaodeng Jiaoyu Chubanshe
Issue Date: 2011
ISSN: 1674-800X
Statement of
Benjamin Rowlatt, Jake A. Yorke, Anthony J. Strong, Christopher J. C. Whitehouse, Stephen G. Bell, Luet-Lok Wong
Abstract: Fatty acid binding and oxidation kinetics for wild type P450(sub)BM3 (CYP102A1) from Bacillus megaterium have been found to display chain length-dependent homotropic behavior. Laurate and 13-methyl-myristate display Michaelis-Menten behavior while there are slight deviations with myristate at low ionic strengths. Palmitate shows Michaelis-Menten kinetics and hyperbolic binding behavior in 100 mmol/L phosphate, pH 7.4, but sigmoidal kinetics (with an apparent intercept) in low ionic strength buffers and at physiological phosphate concentrations. In low ionic strength buffers both the heme domain and the full-length enzyme show complex palmitate binding behavior that indicates a minimum of four fatty acid binding sites, with high cooperativity for the binding of the fourth palmitate molecule, and the full-length enzyme showing tighter palmitate binding than the heme domain. The first flavin-to-heme electron transfer is faster for laurate, myristate and palmitate in 100 mmol/L phosphate than in 50mmol/L Tris (pH 7.4), yet each substrate induces similar high-spin heme content. For palmitate in low phosphate buffer concentrations, the rate constant of the first electron transfer is much larger than k(sub)cat. The results suggest that phosphate has a specific effect in promoting the first electron transfer step, and that P450(sub)BM3 could modulate Bacillus membrane morphology and fluidity via palmitate oxidation in response to the external phosphate concentration.
Keywords: P450BM3; monooxygenase; fatty acid; cooperativity; allosteric effect; CYP102A1
Rights: © Higher Education Press and Springer-Verlag Berlin Heidelberg 2011
RMID: 0020121227
DOI: 10.1007/s13238-011-1082-6
Appears in Collections:Chemistry and Physics publications

Files in This Item:
There are no files associated with this item.

Items in DSpace are protected by copyright, with all rights reserved, unless otherwise indicated.