Grant: Research Tools and Instruments

In the analysis of essential biological molecules, such as proteins and polysaccharides, size is a critical piece of fundamental information. With proteins, the ability to form homooligomers or heterooligomers (i.e. with other partners), or to interact with ligands, are properties that are invariably essential for function. For polysaccharides, size generally determines their functional properties in biological systems, or their applications as commercial products. While simple in concept, measuring the size of biological macromolecules by size exclusion chromatography is complicated by factors including interaction of the macromolecule with the column matrix, a conformation that may be quite different to available standards, oligomerization or aggregation, and heterogeneity in the sample. These limitations can be overcome by integrating size exclusion chromatography and multi-angle light scattering (SEC-MALS). The requested OMNISEC system includes a dedicated liquid chromatography platform integrated with a 20-angle light scattering detector, as well as photodiode, refractive index detector, and viscometry detectors integrating data from a wide range of biophysical parameters. This systemaffords an absolute measurement of molar mass and rms radius (r g ) for macromolecules in solution and (to the best of our knowledge) would be the only one of its kind at Guelph. It will be housed in space dedicated to protein crystallization and analysis, with secure access limited to those with the appropriate training.The named investigators will use the SEC-MALS system as an integral part of their workflows to examine the structures and functions of enzymes, transporters, and structural proteins involved in bacterial cell-surface assembly, as well as proteins that form microcompartments for protected metabolism in bacteria. However, the additional capacity will be available to other researchers in the Department involved in studying the structure and functions of proteins from diverse biological systems including bacteria, fungi, plants and mammalian cells. The instrument will be an important contributor to leadership and International competitiveness in these research groups. It will also have a major impact on HQP training. The applicants' research groups include 26 current HQP but, with other users, we anticipate this instrument advancing the research efforts of more than 50 HQP, at levels ranging from research associates through to undergraduates. As many of our trainees move to research environments in academia and in biotech-pharma, experience with leading contemporary approaches is an essential part of their marketable skills. The acquisition of the SEC-MALS system will contribute to their leadership and international competitiveness.