Spectrofluorimeter for protein and peptide research

用于蛋白质和肽研究的荧光分光光度计

• 批准号: RTI-2023-00523
• 负责人: Aluko, Rotimi
• 金额: $ 3.16万
• 依托单位: University of Manitoba
• 依托单位国家: 加拿大
• 项目类别: Research Tools and Instruments
• 财政年份: 2022
• 资助国家: 加拿大
• 起止时间: 2022-01-01 至 2023-12-31
• 项目状态: 已结题
• 来源: https://www.nserc-crsng.gc.ca/ase-oro/Details-Detailles_eng.asp?id=753490
• 关键词: Spectrofluorimeter; protein; peptide; research

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.

蛋白质和多肽以及酶-肽复合物的三维构象和折叠构象的测定是蛋白质结构-功能评价和发现新的生物活性肽的关键要求。为了进行这些结构研究，需要测量蛋白质和肽的固有荧光，因为所提供的信息定义了微观和宏观环境，从而能够在各种实验条件下准确解释蛋白质和肽的三维结构。荧光分光光度计是唯一能够提供内在荧光数据的仪器，因此是结构-功能研究所需基础设施的关键组成部分。在马尼托巴大学食品与人类营养科学系，只有一台荧光计，这是我在2005年通过NSERC RTI赠款购买的。然而，这台使用了17年的仪器已经不能使用了，因为该型号已经停产，而且马尼托巴大学校园里没有合适的替代系统。因此，本提案的目的是用新的荧光计取代不可使用的17年系统，这将为我的NSERC和加拿大研究主席资助的生物活性肽研究项目的持续运作提供关键支持，并加强行业研究合作。该仪器需要在存在和不存在生物活性肽的情况下测量酶的构象。对酶-肽复合物性质的准确分析使得能够鉴定肽上提供酶活性的有效抑制的氨基酸残基。这很重要，因为我的NSERC资助的发现计划专注于阐明新的食物蛋白衍生生物活性肽的结构-功能特性。由于酶-肽相互作用对分子结构的影响代表了定义肽功能的关键步骤，因此荧光计是我的Discovery研究工作和成功培训高素质人员（HQP）的必要组成部分。该荧光计将用于阐明生物活性肽的氨基酸序列与其与靶酶的结合亲和力之间的关系。荧光计的输出将提高蛋白质作物（生物活性肽生产的底物）的增值利用率，并加强加拿大食品和营养行业的经济价值。荧光计将被研究生和本科生，博士后研究员/研究助理和技术人员每天用于研究酶-肽复合物的结构特性，这可能导致发现新的生物活性肽。因此，该工具对于成功培训HQP，为他们提供有助于加拿大食品和生物技术行业全球竞争力的市场技能至关重要。