Please use this identifier to cite or link to this item: http://hdl.handle.net/2440/124619
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Type: Journal article
Title: Structural insights into the role of the acid-alcohol pair of residues required for dioxygen activation in cytochrome P450 enzymes
Author: Coleman, T.
Stok, J.E.
Podgorski, M.N.
Bruning, J.B.
De Voss, J.J.
Bell, S.G.
Citation: JBIC Journal of Biological Inorganic Chemistry, 2020; 25(4):583-596
Publisher: Springer Nature
Issue Date: 2020
ISSN: 0949-8257
1432-1327
Statement of
Responsibility: 
Tom Coleman, Jeanette E. Stok, Matthew N. Podgorski, John B. Bruning, James J. De Voss, Stephen G. Bell
Abstract: The cytochrome P450 heme monooxygenases commonly use an acid-alcohol pair of residues, within the I-helix, to activate iron-bound dioxygen. This work aims to clarify conflicting reports on the importance of the alcohol functionality in this process. Mutants of the P450, CYP199A4 (CYP199A4D251N and CYP199A4T252A), were prepared, characterised and their crystal structures were solved. The acid residue of CYP199A4 is not part of a salt bridge network, a key feature of paradigmatic model system P450cam. Instead, there is a direct proton delivery network, via a chain of water molecules, extending to the surface. Nevertheless, CYP199A4D251N dramatically reduced the activity of the enzyme consistent with a role in proton delivery. CYP199A4T252A decreased the coupling efficiency of the enzyme with a concomitant increase in the hydrogen peroxide uncoupling pathway. However, the effect of this mutation was much less pronounced than reported with P450cam. Its crystal structures revealed fewer changes at the I-helix, compared to the P450cam system. The structural changes observed within the I-helix of P450cam during oxygen activation do not seem to be required in this P450. These differences are due to the presence of a second threonine residue at position 253, which is absent in P450cam. This threonine forms part of the hydrogen bonding network, resulting in subtle structural changes and is also present across the majority of the P450 superfamily. Overall, the results suggest that while the acid-alcohol pair is important for dioxygen activation this process and the method of proton delivery can differ across P450s.
Keywords: Metalloenzymes; heme; cytochrome P450; mechanism; x-ray crystallography; dioxygen activation
Rights: © Society for Biological Inorganic Chemistry (SBIC) 2020
RMID: 1000018341
DOI: 10.1007/s00775-020-01781-4
Grant ID: http://purl.org/au-research/grants/arc/DP140103229
http://purl.org/au-research/grants/arc/FT140100355
Appears in Collections:Chemistry publications

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