1ZO4

Crystal Structure Of A328S Mutant Of The Heme Domain Of P450BM-3

Summary for 1ZO4

• Entry DOI: 10.2210/pdb1zo4/pdb
• Related: 1BU7 1JPZ 1ZO9 1ZOA 2HPD
• Descriptor: Bifunctional P-450:NADPH-P450 reductase, PROTOPORPHYRIN IX CONTAINING FE, 2-(N-MORPHOLINO)-ETHANESULFONIC ACID, ... (5 entities in total)
• Functional Keywords: cytochrome p-450, hemeprotein a328s, oxidoreductase
• Biological source: Bacillus megaterium
• Cellular location: Cytoplasm (By similarity): P14779
• Total number of polymer chains: 2
• Total formula weight: 111294.17

Authors

Hegda, A.,Chen, B.,Haines, D.C.,Bondlela, M.,Mullin, D.,Graham, S.E.,Tomchick, D.R.,Machius, M.,Peterson, J.A. (deposition date: 2005-05-12, release date: 2006-08-01, Last modification date: 2023-08-23)

Primary citation

Haines, D.C.,Hegde, A.,Chen, B.,Zhao, W.,Bondlela, M.,Humphreys, J.M.,Mullin, D.A.,Tomchick, D.R.,Machius, M.,Peterson, J.A.
A single active-site mutation of P450BM-3 dramatically enhances substrate binding and rate of product formation.
Biochemistry, 50:8333-8341, 2011

PubMed Abstract: Identifying key structural features of cytochromes P450 is critical in understanding the catalytic mechanism of these important drug-metabolizing enzymes. Cytochrome P450BM-3 (BM-3), a structural and mechanistic P450 model, catalyzes the regio- and stereoselective hydroxylation of fatty acids. Recent work has demonstrated the importance of water in the mechanism of BM-3, and site-specific mutagenesis has helped to elucidate mechanisms of substrate recognition, binding, and product formation. One of the amino acids identified as playing a key role in the active site of BM-3 is alanine 328, which is located in the loop between the K helix and β 1-4. In the A328V BM-3 mutant, substrate affinity increases 5-10-fold and the turnover number increases 2-8-fold compared to wild-type enzyme. Unlike wild-type enzyme, this mutant is purified from E. coli with endogenous substrate bound due to the higher binding affinity. Close examination of the crystal structures of the substrate-bound native and A328V mutant BMPs indicates that the positioning of the substrate is essentially identical in the two forms of the enzyme, with the two valine methyl groups occupying voids present in the active site of the wild-type substrate-bound structure.

PubMed: 21875028
DOI: 10.1021/bi201099j

Experimental method

X-RAY DIFFRACTION (1.46 Å)