AUTOMATED CRYSTALLIZATION SCREENING OF REDOX ACTIVE PROTEINS

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This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. Primary support for the subproject and the subproject's principal investigator may have been provided by other sources, including other NIH sources. The Total Cost listed for the subproject likely represents the estimated amount of Center infrastructure utilized by the subproject, not direct funding provided by the NCRR grant to the subproject or subproject staff. Advances in automated high throughput liquid handling have resulted in dramatically reduced sample requirement for macromolecular crystallization. Commercially available crystallization robots, such as the Gryphon produced by Art Robbins Inc, allow for nanoliter drop sizes and thus can screen for conditions that produce X-ray diffraction-quality crystals from as little as 1-2 mg of purified protein. Because the requirement for large amounts of sample is a major impediment to collaborative structural studies, reducing sample needs will expand the scope and quantity of collaborative structural work that can be pursued. This proposal requests funds to acquire the Gryphon crystallization robot. We propose that this acquisition will allow for greatly enhanced structural biological collaboration within the Redox Biology Center and the larger university community. Two RBC crystallographers, Drs. Joe Barycki and Mark Wilson, will be responsible for the instrument and for managing collaborations. Funds are also requested for a support person at the B.S. to M.S. level to maintain the instrument and to handle operational details of these collaborative efforts.