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Tryptophan-96 in cytochrome P450 BM3 plays a key role in enzyme survival

Ravanfar, Raheleh and Sheng, Yuling and Gray, Harry B. and Winkler, Jay R. (2023) Tryptophan-96 in cytochrome P450 BM3 plays a key role in enzyme survival. FEBS Letters, 597 (1). pp. 59-64. ISSN 0014-5793. doi:10.1002/1873-3468.14514. https://resolver.caltech.edu/CaltechAUTHORS:20221107-999291600.27

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Abstract

Flavocytochrome P450 from Bacillus megaterium (P450_(BM3)) is a natural fusion protein containing reductase and hame domains. In the presence of NADPH and dioxygen the enzyme catalyses the hydroxylation of long-chain fatty acids. Analysis of the P450BM3 structure reveals chains of closely spaced tryptophan and tyrosine residues that might serve as pathways for high-potential oxidizing equivalents to escape from the hame active site when substrate oxidation is not possible. Our investigations of the total number of enzyme turnovers before deactivation have revealed that replacement of selected tryptophan and tyrosine residues with redox inactive groups leads to a twofold reduction in enzyme survival time. Tryptophan-96 is critical for prolonging enzyme activity, suggesting a key protective role for this residue.


Item Type:Article
Related URLs:
URLURL TypeDescription
https://doi.org/10.1002/1873-3468.14514DOIArticle
ORCID:
AuthorORCID
Ravanfar, Raheleh0000-0003-2992-0575
Gray, Harry B.0000-0002-7937-7876
Winkler, Jay R.0000-0002-4453-9716
Additional Information:This research was supported by the National Institute of Diabetes and Digestive and Kidney Diseases of the National Institutes of Health under Award Number R01DK019038. The content is solely the responsibility of the authors and does not necessarily represent the official views of the National Institutes of Health. Additional support was provided by the Arnold and Mabel Beckman Foundation.
Funders:
Funding AgencyGrant Number
NIHR01DK019038
Arnold and Mabel Beckman FoundationUNSPECIFIED
Issue or Number:1
DOI:10.1002/1873-3468.14514
Record Number:CaltechAUTHORS:20221107-999291600.27
Persistent URL:https://resolver.caltech.edu/CaltechAUTHORS:20221107-999291600.27
Usage Policy:No commercial reproduction, distribution, display or performance rights in this work are provided.
ID Code:117755
Collection:CaltechAUTHORS
Deposited By: Research Services Depository
Deposited On:18 Nov 2022 21:26
Last Modified:25 Jan 2023 20:42

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