5JQV

Crystal structure of Cytochrome P450 BM3 heme domain T269V/L272W/L322I/A406S (WIVS) variant with iron(III) deuteroporphyrin IX bound

Summary for 5JQV

• Entry DOI: 10.2210/pdb5jqv/pdb
• Related: 5JQU
• Descriptor: Bifunctional cytochrome P450/NADPH--P450 reductase, FE(III) DEUTEROPORPHYRIN IX (3 entities in total)
• Functional Keywords: p450 bm3, iron(iii) deuteroporphyrin ix, oxidoreductase
• Biological source: Bacillus megaterium (strain ATCC 14581 / DSM 32 / JCM 2506 / NBRC 15308 / NCIMB 9376 / NCTC 10342 / VKM B-512)
• Total number of polymer chains: 8
• Total formula weight: 436710.74

Authors

Reynolds, E.W.,McHenry, M.W.,Cannac, F.,Gober, J.G.,Snow, C.D.,Brustad, E.M. (deposition date: 2016-05-05, release date: 2016-09-28, Last modification date: 2023-09-27)

Primary citation

Reynolds, E.W.,McHenry, M.W.,Cannac, F.,Gober, J.G.,Snow, C.D.,Brustad, E.M.
An Evolved Orthogonal Enzyme/Cofactor Pair.
J.Am.Chem.Soc., 138:12451-12458, 2016

PubMed Abstract: We introduce a strategy that expands the functionality of hemoproteins through orthogonal enzyme/heme pairs. By exploiting the ability of a natural heme transport protein, ChuA, to promiscuously import heme derivatives, we have evolved a cytochrome P450 (P450BM3) that selectively incorporates a nonproteinogenic cofactor, iron deuteroporphyrin IX (Fe-DPIX), even in the presence of endogenous heme. Crystal structures show that selectivity gains are due to mutations that introduce steric clash with the heme vinyl groups while providing a complementary binding surface for the smaller Fe-DPIX cofactor. Furthermore, the evolved orthogonal enzyme/cofactor pair is active in non-natural carbenoid-mediated olefin cyclopropanation. This methodology for the generation of orthogonal enzyme/cofactor pairs promises to expand cofactor diversity in artificial metalloenzymes.

PubMed: 27575374
DOI: 10.1021/jacs.6b05847

Experimental method

X-RAY DIFFRACTION (2.34 Å)