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Structure-Guided Recombination Creates an Artificial Family of Cytochromes P450

Otey, Christopher R. and Landwehr, Marco and Endelman, Jeffrey B. and Hiraga, Kaori and Bloom, Jesse D. and Arnold, Frances H. (2006) Structure-Guided Recombination Creates an Artificial Family of Cytochromes P450. PLoS Biology, 4 (5). e112. ISSN 1544-9173. PMCID PMC1431580. http://resolver.caltech.edu/CaltechAUTHORS:OTEpbio06

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Use this Persistent URL to link to this item: http://resolver.caltech.edu/CaltechAUTHORS:OTEpbio06

Abstract

Creating artificial protein families affords new opportunities to explore the determinants of structure and biological function free from many of the constraints of natural selection. We have created an artificial family comprising ~3,000 P450 heme proteins that correctly fold and incorporate a heme cofactor by recombining three cytochromes P450 at seven crossover locations chosen to minimize structural disruption. Members of this protein family differ from any known sequence at an average of 72 and by as many as 109 amino acids. Most (>73%) of the properly folded chimeric P450 heme proteins are catalytically active peroxygenases; some are more thermostable than the parent proteins. A multiple sequence alignment of 955 chimeras, including both folded and not, is a valuable resource for sequence-structure-function studies. Logistic regression analysis of the multiple sequence alignment identifies key structural contributions to cytochrome P450 heme incorporation and peroxygenase activity and suggests possible structural differences between parents CYP102A1 and CYP102A2.


Item Type:Article
Related URLs:
URLURL TypeDescription
https://www.ncbi.nlm.nih.gov/pmc/articles/PMC1431580/PubMed CentralArticle
https://doi.org/10.1371/journal.pbio.0040112DOIArticle
ORCID:
AuthorORCID
Bloom, Jesse D.0000-0003-1267-3408
Arnold, Frances H.0000-0002-4027-364X
Additional Information:© 2006 Otey et al. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited. The NCBI Entrez (http://www.ncbi.nlm.nih.gov/gquery/gquery.fcgi) accession numbers for the genes and gene products discussed in this paper are CYP102A1 (J04832), CYP102A2 (CAB12544) and CYP102A3 (U93874). We thank Kiowa S. Bower for laboratory assistance. CRO, ML, KH, and FHA conceived and designed the experiments. CRO, ML, and KH performed the experiments. CRO, ML, JBE, JDB, and FHA analyzed the data. JBE, JDB, and FHA contributed reagents/materials/analysis tools. CRO, ML, and FHA wrote the paper. The authors have declared that no competing interests exist.
Issue or Number:5
PubMed Central ID:PMC1431580
Record Number:CaltechAUTHORS:OTEpbio06
Persistent URL:http://resolver.caltech.edu/CaltechAUTHORS:OTEpbio06
Alternative URL:http://biology.plosjournals.org/perlserv/?request=get-document&doi=10.1371/journal.pbio.0040112
Official Citation:Otey CR, Landwehr M, Endelman JB, Hiraga K, Bloom JD, et al. (2006) Structure-Guided Recombination Creates an Artificial Family of Cytochromes P450. PLoS Biol 4(5): e112
Usage Policy:No commercial reproduction, distribution, display or performance rights in this work are provided.
ID Code:2699
Collection:CaltechAUTHORS
Deposited By: Archive Administrator
Deposited On:20 Apr 2006
Last Modified:21 Aug 2018 20:15

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