A liquid chromatography system with integrated multi-angle light scattering detection to characterize protein folding, assembly, and modification

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This application requests funds to purchase a multi-angle light scattering detector coupled to a suitable size exclusion chromatography system (SEC-MALS). SEC-MALS provides a direct measurement of the molecular mass and radius of gyration of biomacromolecules. It also allows for molecules to be characterized during the purification process, which is essential for unstable and time-sensitive samples. These capabilities do not exist in Atlantic Canada, and are urgently needed to enhance the applicants' research capabilities. All five co-applicants and their 23 current trainees, ranging from undergraduate student to postdoctoral fellow, will immediately take advantage of this instrument for the study of fundamental processes of life and biomaterial development. SEC-MALS is a gold-standard technique to determine protein purity and homogeneity, and will be routinely used by all of the applicants to monitor the quality of their research samples. Acquisition of a SEC-MALS instrument will also enable new directions of study that investigate the self-assembly of silk, filamentous enzymes, and fungal proteins, where these proteins are often in a dynamic equilibrium involving several oligomeric states. Other research projects will rely upon the benefits of SEC-MALS to efficiently detect and quantify protein modification by intein splicing and ubiquitination, as well as monitoring protein-protein interactions involving transcription factors. This instrument will be installed in the Biophysical Analysis Facility at Dalhousie, providing unfettered access to all of the co-applicants, their trainees, and to other researchers in the Halifax region. This instrument will be operated directly by students and other highly qualified personnel as a part of their research training, each of whom will be mentored in hands-on use of the instrumentation and in expert analysis of the resulting data.