Development of a Novel Test System for Screening Antagonists of ErbB Receptors in Breast Cancer
Final rept. 1 Jun 2000-31 May 2004
STANFORD UNIV CA
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The focus of this project was to design a system for screening chemical compounds that would inhibit the dimerization of the EGFR family members ErbB2 and ErbB3 using beta-galactosidase complementation. We attempted to apply the beta galactosidase complementation system to both ErbB2, 3 homo and heterodimerization, as well as the interaction of these receptors with downstream signaling molecules such as Grb2. The initial attempts to design an assay amenable to high-throughput screening for antagonists of ErbB2 activity failed due to fundamental problems associated with the interaction system employed. In order to circumvent these limitations we rationally redesigned the system to be more stable and tolerate wider ranges of expression levels. The new system based on a 46aa mutated alpha-peptide was able to successfully discern inducible interactions for transmembrane receptors, cytosolic, and nuclear proteins. Further the magnitude of induction is sufficient for high throughput screening for drugs that inhibit this heterodimerization event.
- Medicine and Medical Research
- Polymer Chemistry