Bioorganic activation of cytochrome P-450cam

The Cytochrome P-450 class of monoxygenases carry out a wide variety of hydroxylation, epoxidation and heteroatom oxidation reactions. The cytochrome P-450cam enzyme from P. putida has been widely studied as a model for other P-450s due to the availability of a high resolution X-ray crystal structure. In this thesis several questions concerning the activation of P-450cam by substrate, electron transfer and oxygen cleavage are addressed. The identity of the cysteine ligands to the (2Fe-2S) center of P-450cam's electron transport partner, putidaredoxin (Pdx), is determined using a combination of site-directed mutagenesis and EPR spectroscopy. The identity of the rapidly reacting cysteine in P-450cam, which has been used as an attachment site for probes of Pdx-P450cam interaction, is identified. The roles of several amino acids in and around the P-450cam active site are studied in order to better understand their action in substrate access and binding. To help determine the location of the substrate access channel in P-450cam, a novel disulfide bridge and a site for an intraprotein chemical crosslink have been engineered and characterized.

Name thesis:degree_name

Ph.D.

Level thesis:degree_level

Dissertation

Discipline thesis:degree_discipline

Biochemistry

Grantor

University of Illinois at Urbana-Champaign

Year dc:date

2011

Author dc:creator

• Gerber, Nancy Counts

Contributors dc:contributor

• Sligar, Stephen G.

• Chemistry, Biochemistry

Statement dc:rights

• Copyright 1993 Gerber, Nancy Counts

Language dc:language

eng

Identifier

AAI9411631
(UMI)AAI9411631

OAI identifier oai:identifier

oai:www.ideals.illinois.edu:2142/20647