Training in Support of Research Project Entitled "Tumor Cell De-adhesion by Aberrant, Single Skubmit Integrins".
Abstract
My purpose is try to identify a plasma membrane protein fingerprint which can be used to distinguish invasive breast cancer cell lines from non-invasive breast cancer lines. The methods included those for purification of plasma membrane proteins , extraction of proteins using urea/thiourea mixture, a standard procedure for 2D PAGE, excision of polypeptide "spots" from gels, trypsin digestion, MALDI TOF mass spectrometry and database searching for protein identity are well-established for this project. Three breast cancer cell lines (MDA-MB435, MDA-MB23 1 and MCF7) with different metastatic potential have been grown and 2-D maps of plasma membrane proteins have been generated in the laboratory. Computer- assisted analysis of the 2D polypeptide pattern showed that 56% of the polypeptide spots were the same between MCF7 and MDA-MB23 1 (two non-metastatic Breast Cancer cell lines), only 42% of the polypeptide spots were the same between MCF7 and MDA-MB435 (metastatic vs non-metastatic Breast Cancer cell lines). This differential expression of proteins is being characterized in order to determine which proteins are important to the metastatic condition. A number of proteins from the 2D gels has been identified by MALDI TOF mass spectrometry. Recently, antibody library phage display, has been expanded for further identifying the plasma membrane proteins.
Document Details
- Document Type
- Technical Report
- Publication Date
- Jun 01, 1999
- Accession Number
- ADA379615
Entities
People
- Yan Zhang
Organizations
- Sanford Burnham Prebys Medical Discovery Institute