PROTEIN CRYSTALLOGRAPHY FOR BIOCHEMISTS

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Protein structure solution is now a required step in many important fields from molecular biology, enzymology to drug design. Protein crystallography is still the most important method for this step. One exciting new development of protein crystallography is that it can be made so routine that a biochemist group with the help of a protein crystallographer could begin to crystallize and solve protein structures. This development is of great significance because biochemists usually have the most interesting proteins. It is, therefore, very important to set up a Protein Crystallography Data Collection Facility that would be shared by experienced protein crystallographers and biochemist. This facility would serve two purposes: One would be to provide many research groups with the most modern methods and instruments to collect data, and the other would be to provide an opportunity to enhance the contacts between crystallographers and biochemists. These contacts are crucial for biochemists in order to avoid publishing wrong structures. The aim of this proposal is to buy equipment to set up this facility. This data collection facility will include a MAR Imaging Plate detector, a set of large focusing mirrors, a cryo device and an on line computer. This system will be set up on a RU200 X-ray generator already available in the P.I's laboratory. This data collection facility will be shared by 13 research groups: 3 experienced protein crystallographer groups, and 10 biochemist/biophyscist groups. The research activities of these groups are many fold - from solving the structure of proteins involved in transduction signal-like cAMP dependent protein kinase, to the examination of structures of whole viruses. Other interesting proteins that will be studied are fibrinogens, Reaction Center, F1 -ATPase, dihydropteridine reductase, thrombomodulin complexes, virulence factors of bacterial pathogens, different transcription factors etc. Another very important function of this facility will be the training of future protein crystallographers. This facility will be used primarily by undergraduate and graduate students and postdoctoral fellows. Coupling to the activities of this facility will be a course in Protein Crystallography that will be taught by the Principal Investigator. Another course on Macromolecular Modeling is being offered by the Chemistry & Biochemistry department in collaboration with the San Diego Supercomputer Center. In addition, we plan to offer, every year, a three-day workshop on protein crystallization methods for noncrystallographers.