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CSB Special Seminar: Dr. Ivan Shabalin, Dept of Molecular Physiology & Biological Physics, University of Virginia, Charlottesville, VA
June 17, 2016 @ 11:00 am - 12:00 pm
Cell & Systems Biology
Special Seminar
Dr. Ivan Shabalin
Research Scientist
Dept of Molecular Physiology & Biological Physics
University of Virginia
Charlottesville, VA
“Transformation of High-Throughput Protein Crystallization Screening into High-Output”
Abstract:
Many protein crystallization methods and tools have been developed over the past decades. Among them are many commercially available screens and numerous protein modification methods for improvement of protein “crystallizability,” such as affinity tag removal/retention, methylation, surface mutagenesis and limited proteolysis. Given the options, choosing an effective crystallization strategy can be a daunting task, especially if the supply of protein is limited and protein production is expensive. In addition, many of those methods are relatively time-consuming, and require significant amounts of consumables. The use of the LabDB/XtalDB database system allowed us to systematically study the crystallization of a large number of protein targets and perform comparative analysis of different methods and approaches. In our recent work on a set of 97 NYSGRC targets, we used various techniques to develop an optimized strategy for protein crystallization, which resulted in the determination of the structures of 42 targets (a 43% success rate). The study was guided by multiple goals—maximizing the number of successful crystallizations, yet limiting the resources necessary for screening and optimization by including tests of some less popular, yet promising, techniques. The methods we used included initial screening with alternative reservoirs, limited proteolysis in situ, co-crystallization with function-based potential ligands, “air-dry” cryoprotection of crystals, and many others. Based on our experience, we suggest an easy-to-follow protein crystallization strategy, which is cost- and time-effective but has proved to be high-output rather than high-throughput.
If time permits, he will also discuss a second topic: Reliability of protein crystal structures for structure-based drug discovery.