Differential Scanning Calorimeter for Protein Studies

Differential scanning calorimetry (DSC) is a powerful experimental technique for determining the thermodynamic properties accompanying protein-protein binding, protein unfolding, and ligand binding. This grant will provide a differential scanning calorimeter, allowing new experiments in protein characterization to be conducted at Dalhousie University. The ability of DSC to monitor the unfolding, phase transitions, and degree of oligomerization of proteins, polynucleotides, and lipid assemblies makes it extremely useful for a variety of applications. All 6 NSERC-funded applicants and their 42 trainees, ranging from undergraduates to postdoctoral fellows and professional research associates, with interests spanning the life and biomedical sciences to fundamental and applied materials chemistry, will immediately be able to take advantage of this valuable biophysical technique. The instrument will permit optimum calorimetric characterization of proteins such as spider silks, viral proteins, antifreeze proteins, collagen, and enzymes, with the potential for new biomedical or biotechnological applications. Specifically, applicants and trainees will be able to (1) evaluate the energetic contributions of noncovalent interactions to the stabilization of enzyme-ligand adducts, (2) assess the effect of interdigitating loops on the stability of enzyme oligomers, (3) determine the stability of recombinant silk proteins and proteins bearing specific post-translational modifications to inform protein engineering, (4) delineate the role of protein-protein and protein-lipid interactions governing the ability of viral proteins to mediate membrane fusion events, (5) study the pathways of α-synuclein and antifreeze folding and misfolding and the effect of small molecules that modulate these processes, and (6) elucidate the role of collagen in fibril damage. The fundamental information gained using DSC is essential for understanding enthalpic and entropic contributions to protein stability and intermolecular interactions, and for re-engineering and producing new proteins with desirable properties.No instrument with the sensitivity and appropriate configuration for analyzing dilute protein solutions is available to the applicants either at Dalhousie or nearby institutions, making acquisition essential to support and enhance their research and training programs. The instrument will be installed in Dalhousie’s Proteomics Core Facility, providing ready access to the co-applicants and to many other researchers at Dalhousie and surrounding institutions. Rather than simply providing results as a service, the Facility fully supports the development of trainees’ abilities to conduct experiments and interpret the results. Trainees will gain both hands-on experience and valuable expertise with this important biophysical technique for protein characterization.