Structural investigation of a N-his6 variant of cytochrome P450cam (CYP 101A1) by protein NMR studies

Cytochromes P450 are enzymes that add oxygen to organic molecules, to convert them to water-soluble products that can be further metabolized. These oxidative enzymes are ubiquitous (found in animals, plants and microorganisms). We are working on cytochrome P450cam, isolated from a wild soil bacterium. This enzyme catalyzes the oxidation of camphor (an organic molecule that can be used by these bacteria as a sole carbon and energy source) to 5-exo-hydroxycamphor. The enzyme needs oxygen (O2) for this. We discovered an unexpected reduction reaction catalyzed by this enzyme, to give a camphor-reduced product, borneol, that occurs when O2 levels are insufficient for the bacteria to fully digest camphor. This reaction is used by the bacteria to signal that O2 levels are too low to metabolize camphor, which becomes toxic to them if they cannot digest it. I discovered that a variant of P450cam with additional residues, his6-P450cam, does not catalyze the reduction of camphor to borneol. Based on previous work, we hypothesize that, to reduce camphor, the P450cam needs to be in a closed state, as opposed to open states. “TO BE CONT’D”

Faculty Supervisor:

Erika Plettner

Student:

Partner:

Brandeis University

Discipline:

Life Sciences

Sector:

Life Sciences (not health); Biotechnology; Pharmaceuticals

University:

Simon Fraser University

Program: