A protein purification platform for biochemistry and structural biology research.

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The present proposal is prepared to request a complete system to perform fast protein liquid chromatography (FPLC) in oder to establish a complete platform for protein expression and purifcation at UQÀM. FPLC is an experimental technique that allows the very efficient separation, in non-denaturing and low pressure condition, of a complex ensemble of biomolecules in order to purify, characterize, quantify and identify a particular compound of the initial mixture, usually proteins, protein-complexes and nucleic acids. Briefly, this liquid chromatographic technique is supported by the elution of a mobile phase (analyte) through a stationary phase (column) following by the analysis with a detector. This equipment will play a central role for the achievement of our research programs, as we need highly-pure and well-characterized protein preparation for our biological, biochemical or structural investigations. Several research programs that are underway at the Department will be supported by this apparatus. For instance, we will use this FPLC to isolate soluble protein aggregates that are precursors of amyloid fibrils associated with various diseases for (bio)chemical and biological studies. Besides, this system will support the structural study by NMR of the drug-induced long QT syndrome by allowing the purification of fragments of the ether-à-gogo-related gene (hERG) potassium channel that will be previously expressed in bacteria. Other research projects that will be supported with this infrastructure include the understanding of the structure-to-function impact of the glucose-6-phosphate translocase G6PT, the elucidation of the function of ceruloplasmin in the nervous system development and the study of the cardioprotective potential of copper-containing amine-oxidase enzymes. The research projects that will be supported with this requested FPLC will allow a better understanding of the molecular basis of physiological processes in normal and/or disease state and could ultimately lead to innovative therapeutic approaches for different diseases, such as cancer, the drug-induced long QT syndrome, protein misfolding disorders and developmental disorders associated with neuronal disorganization.