5BV5

Structure of CYP119 with T213A and C317H mutations

Summary for 5BV5

• Entry DOI: 10.2210/pdb5bv5/pdb
• Descriptor: Cytochrome P450 119, PHOSPHATE ION, PROTOPORPHYRIN IX CONTAINING FE, ... (5 entities in total)
• Functional Keywords: p450, heme, p420, cytochrome, oxidoreductase
• Biological source: Sulfolobus solfataricus
• Total number of polymer chains: 4
• Total formula weight: 174804.44

Authors

Buller, A.R.,Heel, T.,McIntosh, J.A.,Arnold, F.H. (deposition date: 2015-06-04, release date: 2016-02-03, Last modification date: 2024-03-06)

Primary citation

McIntosh, J.A.,Heel, T.,Buller, A.R.,Chio, L.,Arnold, F.H.
Structural Adaptability Facilitates Histidine Heme Ligation in a Cytochrome P450.
J.Am.Chem.Soc., 137:13861-13865, 2015

PubMed Abstract: Almost all known members of the cytochrome P450 (CYP) 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.

PubMed: 26299431
DOI: 10.1021/jacs.5b07107

Experimental method

X-RAY DIFFRACTION (2.7 Å)