In this proposal, funds are requested to acquire automated crystallization and imaging systems to support biomedical research at Weill Medical College of Cornell University (WMCCU) and the neighboring Rockefeller University (RU). The instrument will directly support 14 NIH funded researchers and 41 NIH funded research grants and serve as a core resource in the well established X-ray crystallography core facility of the WMCCU. Currently, the X-ray crystallography core facility is equipped with excellent capability for in-house crystal screening and data collection, but lacks any resources for crystallization, a bottleneck for crystallographic pursuit. To allow medium throughput and small volume crystallization of multiple constructs and multiple complexes of challenging macromolecular targets and to facilitate initiation of crystallographic studies by members of a broader scientific community, there is a critical need for a crystallization robotic device at WMCCU and RU. The instruments requested include a semi-automatic solution-handling robot capable of setting crystallization trials using as little as 50 nl protein solution per drop and a semi-automatic imaging system for the analysis and documentation of crystallization trials. The instruments will provide capabilities that do not currently exist in the WMCCU as well as the neighboring RU and the Sloan-Kettering Institute. Both the crystallization and the imaging system will be located in the 200C temperature controlled room in the X-ray crystallography core facility and will be made accessible to both participating PIs in this application and other researchers in the tri-institutional (WMCCU, RU and Sloan-Kettering Institute) area.
X-ray crystallography is one of the most powerful methods for determination of macromolecular structures at atomic level. The acquisition of semi-automated crystallization and imaging systems will directly support NIH sponsored research aimed at understanding basic biological processes such as signal transduction, transcription, ligand transport across membranes and bacterial pathogenesis and at understanding human diseases such as cancer, Alzheimer's disease, Parkinson's disease and microbial infection.
