Novel activities and sequence-function relationships of a synthetic P450 family

• Applicant: Professorin Dr. Andrea Rentmeister
• Subject Area: Biological and Biomimetic Chemistry
• Term: from 2007 to 2010
• Project identifier: Deutsche Forschungsgemeinschaft (DFG) - Project number 50704867

Comparative analyses of naturally occurring protein sequences are widely used to obtain information about the relationship between a protein’s sequence and its structure and function. Whereas analyses of sequences that are related by evolution often suffer from limited and biased information available, synthetic protein families offer new opportunities to explore the determinants of structure and biological function free from the constraints of natural selection. A unique synthetic family comprising 6,561 P450 heme proteins was recently constructed in the group of Prof. Arnold by recombination and shown to be simultaneously highly mutated and properly folded [1]. This family of P450 chimeras represents an unprecedented resource to explore the range of new activities that are accessible by recombination as well as sequence-function relationships. I propose to test the activities of these chimeric enzymes on an array of compounds, including ones that are not accepted by any of the parental proteins, in order to determine the substrate profiles exhibited by the chimeras. I will analyze the data using logistic regression, k-means clustering and a novel combination of both in order to build predictive models of chimera activity on a given substrate or a cluster of substrates, respectively. My work aims at understanding the sequence-function relationship of P450 proteins and shall allow to identify protein sequences capable of hydroxylating a desired compound; these predictions will be experimentally verified.

• DFG Programme: Research Fellowships
• International Connection: USA
• Host: Professorin Frances H. Arnold, Ph.D.