Macromolecular recognition in the cytochrome P450(cam) enzyme system

The cytochrome P-450$\sb{\rm cam}$ reaction cycle is a complex set of coordinated chemical transformations requiring precise temporal and spatial control of reactivities. Cytochrome P-450$\sb{\rm cam}$ catalyzes the regio- and stereo-specific hydroxylation of camphor to form 5-exo-hydroxycamphor. The two reducing equivalents required for this reaction are supplied physiologically by putidaredoxin, a Fe$\sb2$S$\sb2$ iron-sulfur protein. The mammalian cytochromes P-450 are also known to interact with cytochrome b$\sb5$, a small redox protein for which a high resolution crystal structure is available. To characterize the molecular cytochrome P-450$\sb{\rm cam}$ binding surface, cytochrome b$\sb5$ was first genetically engineered to afford a fluorescent derivative capable of monitoring its association with cytochrome P-450$\sb{\rm cam}$. The interaction was subsequently computer modeled by looking for van der Waals complementarity and salt bridge formation between the cytochrome b$\sb5$ anionic binding surface and basic residues on the cytochrome P-450$\sb{\rm cam}$ surface. A good fit was found on the proximal surface of nearest approach to the cytochrome P-450$\sb{\rm cam}$ heme prosthetic group.

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

• Stayton, Patrick Sean

Contributors dc:contributor

• Sligar, Stephen G.

• Chemistry, Biochemistry
• Biophysics, General

Statement dc:rights

• Copyright 1989 Stayton, Patrick Sean

Language dc:language

eng

Identifier

AAI9011038
(UMI)AAI9011038

OAI identifier oai:identifier

oai:www.ideals.illinois.edu:2142/20509