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Structural Adaptability Facilitates Histidine Heme Ligation in a Cytochrome P450

McIntosh, John A. and Heel, Thomas and Buller, Andrew R. and Chio, Linda and Arnold, Frances H. (2015) Structural Adaptability Facilitates Histidine Heme Ligation in a Cytochrome P450. Journal of the American Chemical Society, 137 (43). pp. 13861-13865. ISSN 0002-7863. PMCID PMC4635421. doi:10.1021/jacs.5b07107.

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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.

Item Type:Article
Related URLs:
URLURL TypeDescription Information CentralArticle
McIntosh, John A.0000-0002-9487-490X
Buller, Andrew R.0000-0002-9635-4844
Arnold, Frances H.0000-0002-4027-364X
Additional Information:© 2015 American Chemical Society. This is an open access article published under a Creative Commons Attribution (CC-BY) License, which permits unrestricted use, distribution and reproduction in any medium, provided the author and source are cited. Received: July 8, 2015. Publication Date (Web): August 24, 2015. We thank Jon Rittle, Hans Renata, Todd Hyster, Sheel Dodani, Jackson Cahn, Austin Rice, and Angelo Di Bilio for helpful discussions and comments on earlier versions of this manuscript. We would also like to thank Sabine Brinkmann-Chen, Pavle Nikolovski, and Jens Kaiser for assistance with crystallography screening and structure determination. J.A.M. and A.R.B. are supported by Ruth L. Kirschstein NRSA postdoctoral fellowships from the National Institutes of Health (F32GM101792 and F32GM110851, respectively). T.H. is supported by a FWF Schroedinger fellowship (J3327-B21). We also acknowledge support from the National Science Foundation, Office of Chemical, Bioengineering, Environmental and Transport Systems SusChEM Initiative (grant CBET-1403077), the Caltech Molecular Observatory, which is supported by the Gordon and Betty Moore Foundation, the Beckman Institute, and the Sanofi-Aventis Bioengineering Research Program at Caltech.
Funding AgencyGrant Number
NIH Predoctoral FellowshipF32GM101792
NIH Postdoctoral FellowshipF32GM110851
FWF Der WissenschaftsfondsJ3327-B21
Gordon and Betty Moore FoundationUNSPECIFIED
Caltech Beckman InstituteUNSPECIFIED
Caltech Sanofi-Aventis Bioengineering Research ProgramUNSPECIFIED
Issue or Number:43
PubMed Central ID:PMC4635421
Record Number:CaltechAUTHORS:20150928-085007684
Persistent URL:
Official Citation:Structural Adaptability Facilitates Histidine Heme Ligation in a Cytochrome P450 John A. McIntosh, Thomas Heel, Andrew R. Buller, Linda Chio, and Frances H. Arnold Journal of the American Chemical Society 2015 137 (43), 13861-13865 DOI: 10.1021/jacs.5b07107
Usage Policy:No commercial reproduction, distribution, display or performance rights in this work are provided.
ID Code:60562
Deposited By: Tony Diaz
Deposited On:29 Sep 2015 03:23
Last Modified:24 May 2022 22:29

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