Calpain and calcium-mediated proteolytic regulation of cell signalling networks

钙蛋白酶和钙介导的细胞信号网络的蛋白水解调节

• 批准号: RGPIN-2019-04137
• 负责人: Dufour, Antoine
• 金额: $ 2.19万
• 依托单位: University of Calgary
• 依托单位国家: 加拿大
• 项目类别: Discovery Grants Program - Individual
• 财政年份: 2020
• 资助国家: 加拿大
• 起止时间: 2020-01-01 至 2021-12-31
• 项目状态: 已结题
• 来源: https://www.nserc-crsng.gc.ca/ase-oro/Details-Detailles_eng.asp?id=714647
• 关键词: Calpain; calcium; mediated; proteolytic; regulation

Systems-wide investigation of cell signals adds to our understanding of complex biological processes. My unbiased approach of using global analysis guides my research into deciphering key mechanisms of biology. Proteolysis is a key and irreversible modification but remains understudied at a systems-wide level. It is an essential area of study since the cutting of proteins drastically modifies their functions, yet only ~50% of proteases (enzymes that can cut other proteins) have known substrates. My core expertise in systems biology positions me to pioneer transformative proteomics technologies and will elevate Canada's biochemical knowledge of cell signaling and protease biology during Calcium (Ca2+) activation. Vision and long-term objectives My research program focuses on developing system-wide mass spectrometry approaches to understand the contribution of proteolytic regulation to cell signaling. I will contribute to the field in the following areas: 1- Ameliorate proteomics enrichment methods of proteolysis 2- Refine bioinformatics analysis of cell signaling 3- System-wide identification of protease substrates The results of a protease's activity are defined by what it cuts and to better understand these changes, the substrates of a protease need to be identified and validated. Ca2+ is an essential signaling molecule and among the key downstream effectors of Ca2+ signaling are a family of proteins called calpains that are directly activated by changes in Ca2+ levels. Calpains are key proteases and major effectors of signal transduction, migration and cell death through targeted processing yet little is known about what proteins they cut. This proposal will identify the substrates of calpains and how Ca2+ impact cell physiology. My specific objectives for this NSERC discovery grant are: 1- Identify calpains' substrates usingunbiased quantitative proteomics. 2- Integrate the role of Ca2+ with calpain proteolysis using bioinformatics. 3- Validate the proteins cut by calpains to better understand their physiological roles. Innovative and anticipated outcomes Systems biology creates the potential for entirely new avenues of exploration and promotes constant innovation over iterative cycles. Proteolysis and Ca2+ signaling are fundamental processes in biology yet little is known about their inter-dependency. My research program is focused on using cutting edge proteomics and bioinformatic technologies to investigate the regulated signaling cellular networks during Ca2+ activation and tackle these crucial knowledge gaps. This proposal is designed to characterize what proteins are cut during Ca2+ activation and to complement our current knowledge of signal transduction in immune cells.

对细胞信号的全系统研究增强了我们对复杂生物过程的理解。我使用全局分析的公正方法指导我的研究破译生物学的关键机制。蛋白水解是一种关键且不可逆的修饰，但在系统范围内的研究仍然不足。这是一个重要的研究领域，因为蛋白质的切割会极大地改变其功能，但只有约 50% 的蛋白酶（可以切割其他蛋白质的酶）具有已知的底物。我在系统生物学方面的核心专业知识使我能够开拓变革性蛋白质组学技术，并将提升加拿大在钙 (Ca2+) 激活过程中细胞信号传导和蛋白酶生物学的生化知识。 愿景和长期目标 我的研究项目侧重于开发全系统质谱方法，以了解蛋白水解调节对细胞信号传导的贡献。我将在以下领域为该领域做出贡献： 1-改进蛋白水解的蛋白质组学富集方法 2- 完善细胞信号传导的生物信息学分析 3- 蛋白酶底物的全系统鉴定 蛋白酶活性的结果由其切割的内容决定，为了更好地理解这些变化，需要识别和验证蛋白酶的底物。 Ca2+ 是一种重要的信号分子，Ca2+ 信号传导的关键下游效应器之一是称为钙蛋白酶的蛋白质家族，该蛋白质家族可直接被 Ca2+ 水平的变化激活。钙蛋白酶是关键的蛋白酶，也是通过靶向加工进行信号转导、迁移和细胞死亡的主要效应物，但人们对它们切割的蛋白质知之甚少。该提案将确定钙蛋白酶的底物以及 Ca2+ 如何影响细胞生理学。 我获得 NSERC 发现资助的具体目标是： 1- 使用公正的定量蛋白质组学鉴定钙蛋白酶的底物。 2- 使用生物信息学将 Ca2+ 的作用与钙蛋白酶蛋白水解结合起来。 3- 验证钙蛋白酶切割的蛋白质，以更好地了解其生理作用。 创新和预期成果 系统生物学创造了全新探索途径的潜力，并促进迭代周期的持续创新。蛋白水解和 Ca2+ 信号传导是生物学的基本过程，但人们对它们的相互依赖性知之甚少。我的研究项目专注于利用尖端蛋白质组学和生物信息学技术来研究 Ca2+ 激活过程中受调节的信号传导细胞网络，并解决这些关键的知识空白。该提案旨在描述 Ca2+ 激活过程中哪些蛋白质被切割，并补充我们目前对免疫细胞信号转导的了解。