We proposed to purify from human placenta the cytochrome P-450 enzyme which catalyzes the aromatization of androgens to estrogens. The enzyme will be characterized with respect to substate specificity and kinetics, physicochemical properties and partial amino acid sequence. We also proposed to prepare both polyclonal and monoclonal antibodies to the purified protein. In addition, we propose to investigate the interaction of several types of active site-directed inhibitors as probes of the structure and function of the enzyme. Two types of reversible inhibitors -- non-steroidal compounds which mimic the androgen substrates and a new type of bifunctional steroid inhibitor - will be used for mapping the active site geometry. Several types of chemically reactive steroids -- direct alkylators, suicide substrates and photo-affinity reagents - will be used to covalently label the protein in order to identify regions of the protein sequence which comprise the active site. The inhibitors will be of heuristic use for studying details of the molecular events of the aromatization reaction and will have an application in the design of drugs for the treatment of estrogen-dependent cancers.

Agency
National Institute of Health (NIH)
Institute
National Institute of Diabetes and Digestive and Kidney Diseases (NIDDK)
Type
Research Project (R01)
Project #
5R01DK037551-02
Application #
3236533
Study Section
Biochemical Endocrinology Study Section (BCE)
Project Start
1986-12-01
Project End
1990-06-30
Budget Start
1988-07-01
Budget End
1989-06-30
Support Year
2
Fiscal Year
1988
Total Cost
Indirect Cost
Name
University of California Irvine
Department
Type
Schools of Medicine
DUNS #
161202122
City
Irvine
State
CA
Country
United States
Zip Code
92697