Spectrofluorimeter for protein and peptide research

点击翻译按钮获取中文摘要

Determination of the 3-dimensional and folding conformation of proteins and peptides as well as enzyme-peptide complexes is a critical requirement for structure-function evaluation of proteins and the discovery of novel bioactive peptides. In order to perform these structural studies, measurement of the intrinsic fluorescence of proteins and peptides is a requirement because the information provided defines both the micro and macro environments, enabling accurate interpretation of the 3-dimensional structure of proteins and peptides under various experimental conditions. The spectrofluorimeter is the only instrument capable of providing intrinsic fluorescence data and is, therefore a critical component of the infrastructure needed for structure-function studies. At the Department of Food and Human Nutritional Sciences, University of Manitoba, there is only one fluorimeter, which I purchased in 2005 through an NSERC RTI grant. However, the 17-year-old instrument is no longer serviceable because the model has been discontinued and there is no suitable alternative system on the University of Manitoba campuses. Therefore, the aim of this proposal is to replace the non-serviceable 17-year-old system with a new fluorimeter that will provide critical support for continued operation of my NSERC and Canada Research Chair funded research programs on bioactive peptides and also strengthen industry research collaborations. The instrument is needed to measure the conformation of enzymes in the presence and absence of bioactive peptides. Accurate analysis of the properties of enzyme-peptide complexes enables identification of amino acid residues on peptides that provide potent inhibition of enzyme activity. This is important because my NSERC-funded Discovery program focuses on elucidation of the structure-function properties of novel food protein-derived bioactive peptides. Because the consequences to molecular structure of enzyme-peptide interactions represent a critical step in defining peptide function, the fluorimeter is a required component of my Discovery research work and for successful training of highly qualified personnel (HQP). The fluorimeter will be used to elucidate the relationships between amino acid sequence of bioactive peptides and their binding affinity to target enzymes. The output from the fluorimeter will increase value-added utilization of protein crops (substrates for bioactive peptides production) and strengthen the economic value of the Canadian food and nutrition industry. The fluorimeter will be used by graduate and undergraduate students, postdoctoral fellows/research associates, and technicians on a daily basis to study structural properties of enzyme-peptide complexes, which could lead to the discovery of novel bioactive peptides. Therefore, the instrument is critical to the successful training of HQPs to provide them with marketable skills that contribute to the global competitiveness of Canadian food and biotech industries.