Fast Protein Liquid Chromatograhy System

点击翻译按钮获取中文摘要

This is a reapplication for funds to purchase a GE Healthcare FPLC (Fast Protein Liquid Chromatography) system, needed to replace a 13-year-old FPLC containing several vital components that have become defective over the past 3 years. The FPLC is a pivotal piece of equipment in my lab since it is used virtually on a daily basis for the purification and analysis of protein catalysts (enzymes) from plant tissues and cell cultures, as well as from extracts of transgenic E. coli in which wild-type and mutant versions of several enzymes that we have been studying have been overexpressed. Protein purification forms the foundation upon which the majority of my lab's research is built since our long-term goal is to elucidate the molecular, regulatory, and functional properties of key enzymes of plant carbohydrate and phosphate metabolism. Our research helps to deduce how plants modify and control the chemical reactions that constitute metabolism in response to changing environmental and/or developmental parameters. We are particularly interested in post-translational enzyme modifications by phosphorylation, monoubiquitination, and glycosylation since these can play pivotal roles in controlling enzyme activity, subcellular location, protein:protein interactions and/or turnover in response to various extra- or intracellular signals. Our research has significant long-term applications to problems in Canadian agriculture including the: (1) targeted modification of storage oil versus protein levels in oilseeds such as canola or soybean, (2) optimizing plant-based conversion of atmospheric CO2 into renewable energy sources such as biodiesel and ethanol, and (3) development of phosphate-efficient crops, urgently needed to reduce mankind's rampant but inefficient use of non-renewable, unsustainable, and polluting phosphate fertilizers. This research and associated HQP training would be impossible without ready access to a top-notch FPLC system. The requested FPLC system would also allow lab personnel to exploit many new technologies regarding FPLC design, computer control, chromatographic optimization, and data acquisition that have been developed over the past decade.