4KF2

Structure of the P4509 BM3 A82F F87V heme domain

Summary for 4KF2

• Entry DOI: 10.2210/pdb4kf2/pdb
• Related: 4KEW 4KEY 4KF0
• Descriptor: Bifunctional P-450/NADPH-P450 reductase, PROTOPORPHYRIN IX CONTAINING FE, IMIDAZOLE, ... (4 entities in total)
• Functional Keywords: p450, monooxygenase, oxidoreductase
• Biological source: Bacillus megaterium
• Cellular location: Cytoplasm (By similarity): P14779
• Total number of polymer chains: 2
• Total formula weight: 105994.39

Authors

Leys, D. (deposition date: 2013-04-26, release date: 2013-07-10, Last modification date: 2024-02-28)

Primary citation

Butler, C.F.,Peet, C.,Mason, A.E.,Voice, M.W.,Leys, D.,Munro, A.W.
Key Mutations Alter the Cytochrome P450 BM3 Conformational Landscape and Remove Inherent Substrate Bias.
J.Biol.Chem., 288:25387-25399, 2013

PubMed Abstract: Cytochrome P450 monooxygenases (P450s) have enormous potential in the production of oxychemicals, due to their unparalleled regio- and stereoselectivity. The Bacillus megaterium P450 BM3 enzyme is a key model system, with several mutants (many distant from the active site) reported to alter substrate selectivity. It has the highest reported monooxygenase activity of the P450 enzymes, and this catalytic efficiency has inspired protein engineering to enable its exploitation for biotechnologically relevant oxidations with structurally diverse substrates. However, a structural rationale is lacking to explain how these mutations have such effects in the absence of direct change to the active site architecture. Here, we provide the first crystal structures of BM3 mutants in complex with a human drug substrate, the proton pump inhibitor omeprazole. Supported by solution data, these structures reveal how mutation alters the conformational landscape and decreases the free energy barrier for transition to the substrate-bound state. Our data point to the importance of such "gatekeeper" mutations in enabling major changes in substrate recognition. We further demonstrate that these mutants catalyze the same 5-hydroxylation reaction as performed by human CYP2C19, the major human omeprazole-metabolizing P450 enzyme.

PubMed: 23828198
DOI: 10.1074/jbc.M113.479717

Experimental method

X-RAY DIFFRACTION (1.82 Å)