Lead Inventor:
Eric Gouaux, Ph.D.
Protein identificaiton for crystallography:
Although protein crystallography is a powerful tool in both basic research and drug development, the process of protein crystallization is still poorly understood. This is evidenced by a lack of knowledge about what makes a protein a good candidate for crystallization as opposed to a poor one. Current methods rely on generating large numbers of expression constructs, expressing and purifying the proteins on a large scale, and then characterizing them prior to crystallization. This process is both expensive and time consuming. Ideally, it would be possible to characterize protein constructs prior to large scale expression and purification, reducing both cost and time.
Evaluating proteins as candidates for crystallization experiments:
This invention includes a method for evaluating proteins as candidates for crystallization experiments. The method is consists of performing size exclusion chromatography on the proteins of interest expressing a fluorescent tag. The quantity of protein only needs to be on the order of 10ng and does not need to be in a purified state. As such, the chromatography can be performed on the soluble fraction of total cell lysate. The chromatography is used to detect the fluorescent elution profile of the protein, which can then be characterized to determine likelihood of crystallization. Proteins likely to crystallize have elution profiles that consist of a single, substantially symmetrical peak.
Applications:
• Identification of proteins conducive to crystallization for use in
o Basic scientific research
o Structure-based drug discovery
Advantages:
• Characterize proteins earlier in the crystallization process than current methods
o Decrease costs
o Increase efficiency
• Yields information about a protein's chromatographic behavior
o More predictive of how amenable the protein is to crystallization than the standard yes/no answer for induction and solubility
Patent Status: Patent Pending
Licensing Status: Available for Licensing and Sponsored Research Support
Publications:
Kawate T and Gouaux E (2006). Fluorescence-detection size-exclusion chromatography for precrystallization screening of integral membrane proteins, Structure 14, 673-681.