A Comprehensive Protein Expression, Purification and Crystallization (PEPC) System

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Proteins are the workhorses of all cells, and determining their functions and structures is critical to understanding living organisms. Protein expression and purification allow us to isolate these biological molecules and interrogate their functions and determine their 3-dimensional structures. X-ray crystallography is the most reliable and effective tool to obtain atomic resolution structures of proteins and other biomolecules. Together, structure/function studies contribute to our understanding of biological processes and how these processes might be manipulated for the benefit of manufacturing, food production, agriculture and medicine. Protein expression and purification require large capacity incubators in which to grow bacterial or mammalian cells, and a sophisticated chromatography machine to separate the proteins of interest from other cellular components. Protein crystallization is a laborious, costly and time-consuming process that can be greatly expedited with a crystallization robot. Such infrastructure is essential to a modern biochemistry laboratory. Yet the applicants, and the Molecular Biology and Biochemistry (MBB) department at Simon Fraser University (SFU), lack infrastructure to efficiently express, purify and crystallize proteins, impacting our HQP training, disrupting our research progress and limiting our competitiveness. Funding of this NSERC-RTI proposal will allow the applicants to create a modular, expandable, and cutting-edge Protein Expression, Purification, and Crystallization (PEPC) system that will support biochemistry research at SFU for years to come. This integrated package will include two shaker incubators, a fast liquid phase chromatography instrument, and a crystallization robot, growth chamber and microscope. By modernizing and streamlining the generation of proteins and crystals, the PEPC system will: (1) create substantial labour and reagent cost-savings, (2) significantly enhance HQP training, and (3) expand the research potential of our NSERC-funded programs. The applicants, MBB, and the Faculty of Science at SFU have committed to providing approximately $137K in matching funds, highlighting the need and support for these resources within the community. The proposed PEPC system will serve the diverse and urgent needs of the NSERC-funded applicants. Leading this proposal, the Craig lab will use this infrastructure to solve the structures of Type IV bacterial pilins and other proteins involved in pilus assembly and secretion. The Larijani lab will explore structure/function relationships and novel biochemical properties of DNA/RNA-editing enzymes from evolutionarily distant species. Finally, the Audas lab will examine protein conformational changes associated with functional and pathological amyloid aggregation. Beyond the listed applicants, this resource will be open to all SFU users and will significantly expand our research capacity, productivity and HQP training.