Core B- Expression

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The purpose of this core is to provide cells and proteins as needed to the four Projects of the Program. The specific cells to be isolated by Core B personnel are: 1) blood cells, primarily platelets of human and murine origin; 2) endothelial cells of human and murine origin; and 3) eukaryotic or prokaryotic cells expressing certain integrins, cytoskeletal proteins and mutants of these proteins. The prokaryocytic cells are used as a source for purified proteins; talin, kindlins and scr-family kinase derivatives, in the core and provide to the individual projects as needed. The preparation of cells by experience personnel will help to insure uniformity of purity and function. To have a reliable source of functional platelet preparations as needed by the projects is particularly valuable since the function of platelets can be highly variable. This is also true of endothelial cells of murine origin where experimental progress often depends on performing experiments with the cells derived from a small number of valuable mouse strains. The purified proteins are used in binding studies and for structural analyses, where relatively large quantities of high quality preparations are needed. With the need to prepare a limited number of proteins, isolation procedures can be standardized to provide high quality products. To perform these activities centrally will conserve resources, will help to insure quality and will allow for interproject comparison of results. In addition, since the individual Projects will still require substantial independent cell culture and will undoubtedly need to isolate cells for specific experiments, the Core will function in training members of the Program Project laboratories in proper cell culture and isolation methodologies and in appropriate ways to assess the quality of the cells. Thus, the Core serves an educational function as well.