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加了我们对复杂生物学过程的理解。我使用全球分析的公正方法指导我研究生物学的关键机制。蛋白水解是一种关键且不可逆转的修饰，但仍在全系统层面上进行研究。这是一个重要的研究领域，因为切割蛋白质会大大改变其功能，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将我定位为开创性转化蛋白质组学技术，并将提高加拿大在钙（CA2+）激活过程中对细胞信号传导和蛋白酶生物学的生化知识。 远见和长期目标 我的研究计划着重于开发全系统质谱法的方法，以了解蛋白水解调节对细胞信号的贡献。我将在以下领域为该领域做出贡献： 1-改善蛋白质组学富集蛋白水解方法 2-细胞信号的完善生物信息学分析 3-蛋白酶底物的3-全系统鉴定 蛋白酶活性的结果是由它剪切的内容来定义的，以更好地了解这些变化，需要识别和验证蛋白酶的底物。 Ca2+是一个必不可少的信号分子，在Ca2+信号传导的关键下游效应子中，是一个称为CALPAIN的蛋白质家族，被Ca2+水平的变化直接激活。钙蛋白酶是信号转导，迁移和细胞死亡的主要蛋白酶和主要效应因素，但对它们切割的蛋白质几乎一无所知。该建议将确定CALPAIN的底物以及CA2+如何影响细胞生理。 我对这项NSERC Discovery Grant的具体目标是： 1-使用持续定量蛋白质组学鉴定Calpains的底物。 2-使用生物信息学将Ca2+与钙蛋白酶蛋白水解的作用整合在一起。 3-验证通过钙蛋白酶切割的蛋白质以更好地了解其生理作用。 创新和预期的结果 Systems Biology创造了全新探索途径的潜力，并促进了对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，但对它们的相互依赖性知之甚少。我的研究计划的重点是使用最先进的蛋白质组学和生物信息学技术来研究CA2+激活期间调节的信号细胞网络，并解决这些关键的知识差距。该建议旨在表征在Ca2+激活期间切割的蛋白质，并补充我们当前对免疫细胞信号转导的知识。