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Diversification of Catalytic Function in a Synthetic Family of Chimeric Cytochrome P450s

Landwehr, Marco and Carbone, Martina and Otey, Christopher R. and Li, Yougen and Arnold, Frances H. (2007) Diversification of Catalytic Function in a Synthetic Family of Chimeric Cytochrome P450s. Chemistry and Biology, 14 (3). pp. 269-278. ISSN 1074-5521. PMCID PMC1991292. http://resolver.caltech.edu/CaltechAUTHORS:20180821-082843212

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Abstract

We report initial characterization of a synthetic family of more than 3000 cytochrome P450s made by SCHEMA recombination of 3 bacterial CYP102s. A total of 16 heme domains and their holoenzyme fusions with each of the 3 parental reductase domains were tested for activity on 11 different substrates. The results show that the chimeric enzymes have acquired significant functional diversity, including the ability to accept substrates not accepted by the parent enzymes. K-means clustering analysis of the activity data allowed the enzymes to be classified into five distinct groups based on substrate specificity. The substrates can also be grouped such that one can be a “surrogate” for others in the group. Fusion of a functional chimeric heme domain with a parental reductase domain always reconstituted a functional holoenzyme, indicating that key interdomain interactions are conserved upon reductase swapping.


Item Type:Article
Related URLs:
URLURL TypeDescription
https://doi.org/10.1016/j.chembiol.2007.01.009DOIArticle
https://www.ncbi.nlm.nih.gov/pmc/articles/PMC1991292PubMed CentralArticle
ORCID:
AuthorORCID
Arnold, Frances H.0000-0002-4027-364X
Additional Information:© 2007 Elsevier Ltd. Under an Elsevier user license. Received 20 December 2006, Accepted 22 January 2007, Available online 23 March 2007. This work is supported by the National Institutes of Health (R01 GM068664-0) and a National Science Foundation Predoctoral Fellowship (to M.C.). The authors thank Sally A. Kim for critically reading the manuscript, Daniela C. Dieterich for help with Figure 2, Figure 4, and Christopher Snow for Figure 5. M.L., C.R.O, and F.H.A. designed and planned the project, and M.L., M.C., C.R.O., and Y.L. performed the experiments. M.C., M.L., and C.R.O. analyzed the data. M.L., M.C., and F.H.A. prepared the manuscript.
Funders:
Funding AgencyGrant Number
NIHR01 GM068664-0
NSF Predoctoral FellowshipUNSPECIFIED
Subject Keywords:CHEMBIOL; PROTEINS
PubMed Central ID:PMC1991292
Record Number:CaltechAUTHORS:20180821-082843212
Persistent URL:http://resolver.caltech.edu/CaltechAUTHORS:20180821-082843212
Official Citation:Marco Landwehr, Martina Carbone, Christopher R. Otey, Yougen Li, Frances H. Arnold, Diversification of Catalytic Function in a Synthetic Family of Chimeric Cytochrome P450s, Chemistry & Biology, Volume 14, Issue 3, 2007, Pages 269-278, ISSN 1074-5521, https://doi.org/10.1016/j.chembiol.2007.01.009. (http://www.sciencedirect.com/science/article/pii/S1074552107000373)
Usage Policy:No commercial reproduction, distribution, display or performance rights in this work are provided.
ID Code:88969
Collection:CaltechAUTHORS
Deposited By: Tony Diaz
Deposited On:21 Aug 2018 18:45
Last Modified:21 Aug 2018 18:45

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