Identification of Cellular Binding Sites for a Novel Human Anti-Breast Cancer Peptide
Annual summary rept. 15 Apr 2002-14 Apr 2003
ALBANY MEDICAL COLL NY
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We have synthesized a nine amino acid peptide COP that inhibits the growth of ER human breast. COP does not act like Tam or any other known agent currently used to treat ER breast cancer. Understanding the process by which COP mediates its inhibitory activity is extremely important for the development of this drug or other drugs which exploit the same biochemical pathways to arrest breast cancer growth. A logical first step in this process is to identify the receptor for COP. We have developed and optimized an affinity chromatography procedure to isolate cellular proteins which bind to COP by linking COP to an Affi Gel 10 column. After passing detergent-solubilized cells through the column, retained proteins are sequentially eluted, separated by SDS-PAGE, and visualized by Coomassie staining. To date, a 70 kDa protein has been consistently isolated in this manner, but has not been shown to be specific. Strategies have been identified to address this specificity issue. Binding proteins of interest including the 70 kDa protein described above will be identified and characterized by mass spectrometry, amino acid analysis, and sequence comparison to the known protein databases.
- Medicine and Medical Research