DVLPING SCFV ANTIBODY REAGENTS TO QUANTIFY SIGNAL MOLECULES IN YEAST PHEROMONE

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DESCRIPTION (provided by applicant): Our plan is to establish a P41 Research Resource focused on serving the NIH-supported biomedical research community by developing and integrating new proteomic technologies for collaborative and service studies, disseminating the new technologies, and training scientists in their use. Technological developments within the Resource will contribute significant advances to the efforts of prominent collaborative researchers in the areas of microbial pathogenicity and trauma research, chronic virulent infections, and mammary and melanoma oncology. The ability to precisely measure levels of nearly all expressed proteins and their modified forms can provide new insights into the molecular nature of cells, cell signaling pathways and networks, the cell cycle, cellular differentiation, and other processes relevant to human health and to the progression of various disease states. The ability to characterize protein complexes complements this capability and allows hypotheses to be tested and the biological system operation to be defined. Primary objectives of the Resource are to develop and apply an integrated set of biological methods, new analytical technologies, and associated computational and informatics tools for much more rapid, quantitative, sensitive, and comprehensive proteomic measurements than presently possible; and to facilitate the dissemination of these new technologies to the biomedical research community. Our planned technological developments also aim at providing new capabilities for the characterization of protein modifications and the quantitation of protein abundances spanning more than six orders of magnitude. These fundamental developments will be augmented by 1) capabilities for high throughput isolation of protein complexes needed to obtain better information on protein interactions and to test proteomic hypotheses, and 2) computational and bio-informatics tools needed to effectively extract and visualize data with statistically sound measures of quality to aid in development of new biological understandings. Resource technology developments are directed at challenging proteomic investigations, in collaborations selected on the basis of their scientific and biomedical relevance, as well as their capacity to benefit from the Resource's capabilities and challenge the core technology. The Resource will leverage and extend the substantial capabilities already established in the Environmental Molecular Sciences Laboratory that presently include a collaboratory infrastructure, an integrated and multi-disciplinary scientific team, unique mass spectrometric instrumentation, and a powerful suite of proteome data management and analysis tools.