Critical Replacement of a 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, ligand binding, and the stability of biomolecular assemblies. This grant will replace a malfunctioning, unrepairable differential scanning calorimeter, to permit completion of on-going experiments and publication of the results, and enable new experiments for the characterization of biomacromolecules to be conducted at Dalhousie University. 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. The ability of DSC to monitor the unfolding, phase transitions, and degree of oligomerization of proteins, polynucleotides, lipid assemblies, and other biomolecular structures makes it extremely useful for the study of biomolecules. All 5 NSERC-funded applicants and their 28 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, urgently need to regain access to this valuable biophysical technique. The instrument will permit optimum calorimetric characterization of proteins such as spider silks, antifreeze proteins, hydrophobins, hemolymph proteins, and enzymes, as well as maltodextrin-based particles, with the potential for new biomedical, biotechnological, and ecological applications. Specifically, applicants and trainees will be able to (1) evaluate the role if active site residues to enzyme stability, (2) assess the effect of interdigitating loops on the stability of enzyme oligomers, (3) determine the effects of protein mixing or mutation on the thermal stability of silk fusion proteins and their oligomers, (4) characterize the interactions of antifreeze proteins with cell membranes, (5) develop a novel approach using the hemolymph of blue mussels for environmental monitoring, (6) evaluate the thermal properties of semicrystalline particles used in drug delivery, and (7) assess the stability of assemblies of hydrophobins. 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 Biophysical Characterization Facility, providing ready access to the co-applicants and to many other researchers at Dalhousie and surrounding institutions. The applicants support a diverse and inclusive training environment where trainees will gain both hands-on experience and valuable expertise with this important biophysical technique that is often employed in pharma and biotechnology industries.