CRYSTAL FARM IMAGING SYSTEM: PROTEIN STRUCTURE

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This subproject is one of many research subprojects utilizing the resources provided by a Shared Instrumentation Grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the grant, which is not necessarily the institution for the investigator. DESCRIPTION (provided by applicant): UCSF is in need of state-of-the-art automated instrumentation for crystal management. Our current X-ray facility is used by fourteen independent labs and over seventy people; its user group has doubled over two years, and we anticipate increased demand. About one hundred forty proteins were subjected to crystallization trials in 2004. Automation of crystal storage and retrieval and evaluation of results using network computing is vital. As the number of scientists using the facility increases, efficient use of space and careful monitoring and organization of experiments has become an urgent goal. The DPI CF-400 is a 400- plate storage incubation system that provides users scheduled, high-resolution crystal tray viewing, imaging, scoring, and image storage and tracking. Images of crystals can be taken and archived, all without error- prone human manipulation of crystal trays. As more users vie for limited space, the probability of handling errors is reduced with a device such as the CF-400. Along with scheduled viewing and imaging of crystal trays, access to any plate stored in the incubator is available when a user demands it. The automated crystal imaging system will allow users to remotely determine where slight adjustments to conditions should be made, thus providing faster optimization of crystal-growth conditions and ultimately accelerating all research projects. Barcode plate tracking allows for integration and expandability to accommodate future addition of upstream automated systems, such as nanoliter drop setters. Studies that will benefit from this instrument include ion-channel structure, function, and regulation; kinesins and myosins and their regulatory proteins to learn how molecules transport cellular components; search for novel inhibitors of Eg5, a spindle- associated kinesin, for possible development of anti-cancer agents; Hsp90, known to play a critical role in the regulation of a broad array of proteins involved in intracellular signaling including nuclear steroid receptors such as the estrogen and glucocorticoid receptors; and structure-based design of novel inhibitors of the antibiotic-resistant enzyme beta-lactamase. Relevance to public health: The research involved in this application is focused on the development of anti-cancer agents; understanding the roles and malfunctions of ion channels in areas of the body including the brain; the study of proteins that assist in the folding of steroid receptors; and the design of inhibitors to antibiotic-resistant enzymes.