The Active Site Structure and the Substrate Specificity of Cytochrome P-450(cam)

Cytochrome P-450$\sb{\rm cam}$, from Pseudomonas putida, catalyzes the regio- and stereospecific hydroxylation of the monoterpene d-camphor to afford 5-exo-hydroxycamphor as the sole product. The roles of specific active site structural features in dictating this substrate specificity have been determined with the use of site-directed mutagenesis and specifically modified substrate analogues. In particular, Val-295 forms a complementary fit with the 8,9-gem-dimethyl moiety of camphor and the 10-methyl group of camphor is accommodated by a hydrophobic cleft formed by Leu-244, Phe-98, and Val-247. The site-directed mutant Y96F and the substrate analogue thiocamphor, which perturb the active site hydrogen bond, indicate that the major contribution of this bond is in the maintenance of substrate dependent spin state regulation. Both of these hydrogen bond probes afford a mixed spin system with a decrease in the maximal high spin species obtained.

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

2014

Author dc:creator

• Atkins, William Mark

Contributors dc:contributor

• Sligar, Stephen G.

• Chemistry, Biochemistry

Identifier

(UMI)AAI8908614

OAI identifier oai:identifier

oai:www.ideals.illinois.edu:2142/70574