Tools for High Throughput Protein Crystallization and Structure Determination

DESCRIPTION (provided by applicant): Project Summary: The NIH has committed substantial resources to both traditional and - through its Protein Structure Initiative - high-throughput approaches to protein structure determination, and similar commitments have been made in Europe and Asia. A survey of the results for soluble proteins shows that the main bottlenecks are obtaining high-quality crystals and obtaining high-quality diffraction data for accurate structures. Mitegen, LLC has succeeded in rapidly turning academic discoveries at Cornell into commercial products that are having a major impact on the X-ray diffraction pipeline. Phase I of this project will focus on development of new crystallization platforms based on popular automated drop dispensing technology. Ongoing fundamental and applied studies at Cornell are investigating the physics of protein crystal growth and of the dispensing, dynamics and confinement of protein-containing liquid drops. These studies have led to fundamentally new designs for crystallization plates that promise to reduce storage requirements, allow in-situ X-ray examination of crystal quality, improve equilibration kinetics and the reproducibility of crystallization experiments, and improve the ease with which crystals can be retrieved for X-ray structure determination. Concurrently, tools will be developed for crystal retrieval and X-ray structure determination that integrate with these crystallization platforms and that are suitable for "remote" and automated crystal mounting. These plate and tool designs promise to be highly cost competitive with existing technologies. The Phase I goal is to optimize these designs and begin commercial prototype production. Close collaboration with industrial and academic automation groups (particularly those associated with the PSI) will ensure that the designs are viable. Phase II will focus on more advanced designs incorporating insights from fundamental and applied studies at Cornell, on automated crystal retrieval, and on development of mounts for X-ray microscopies. Relevance: Protein crystallography is a central component of modern structural genomics and drug discovery efforts. By facilitating faster and more efficient protein structure determination, the proposed research will assist in understanding the mechanisms underlying disease and in developing new treatments.