Journal
JOURNAL OF THE AMERICAN CHEMICAL SOCIETY
Volume 137, Issue 43, Pages 13861-13865Publisher
AMER CHEMICAL SOC
DOI: 10.1021/jacs.5b07107
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Funding
- Ruth L. Kirschstein NRSA postdoctoral fellowships from the National Institutes of Health [F32GM101792, F32GM110851]
- FWF Schroedinger fellowship [J3327-B21]
- National Science Foundation
- Office of Chemical, Bioengineering, Environmental and Transport Systems SusChEM Initiative [CBET-1403077]
- Caltech Molecular Observatory
- Gordon and Betty Moore Foundation
- Beckman Institute
- Sanofi-Aventis Bioengineering Research Program at Caltech
- Austrian Science Fund (FWF) [J 3327] Funding Source: researchfish
- Div Of Chem, Bioeng, Env, & Transp Sys
- Directorate For Engineering [1403077] Funding Source: National Science Foundation
- Austrian Science Fund (FWF) [J3327] Funding Source: Austrian Science Fund (FWF)
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Almost all known members of the cytochrome P450 (GYP) superfamily conserve a key cysteine residue that coordinates the heme iron. Although mutation of this residue abolishes monooxygenase activity, recent work has shown that mutation to either serine or histidine unlocks non-natural carbene and nitrene-transfer activities. Here we present the first crystal structure of a histidine-ligated P450. The T213A/C317H variant of the thermostable CYP119 from Sulfolobus acidocaldarius maintains heme iron coordination through the introduced ligand, an interaction that is accompanied by large changes in the overall protein structure. We also find that the axial cysteine C317 may be substituted with any other amino acid without abrogating folding and heme cofactor incorporation. Several of the axial mutants display unusual spectral features, suggesting that they have active sites with unique steric and electronic properties. These novel, highly stable enzyme active sites will be fruitful starting points for investigations of non-natural P450 catalysis and mechanisms.
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