Calpain and calcium-mediated proteolytic regulation of cell signalling networks

对细胞信号的全系统研究增加了我们对复杂生物过程的理解。我不偏不倚的方法使用全局分析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，但在整个系统水平上仍未得到充分研究。这是一个重要的研究领域，因为蛋白质的切割极大地改变了它们的功能，但只有约50%的蛋白酶（可以切割其他蛋白质的酶）有已知的底物。我在系统生物学方面的核心专业知识使我成为转化蛋白质组学技术的先驱，并将提升加拿大在钙（Ca2+）活化过程中细胞信号传导和蛋白酶生物学的生化知识。我的研究计划侧重于开发全系统的质谱方法，以了解蛋白质水解调节对细胞信号传导的贡献。我将在以下领域做出贡献：1-改进蛋白质水解的蛋白质组学富集方法2-改进细胞信号传导的生物信息学分析3-蛋白酶底物的系统范围鉴定蛋白酶活性的结果由它切割的东西来定义，为了更好地理解这些变化，蛋白酶的底物需要被鉴定和验证。Ca2+是一种重要的信号分子，在Ca2+信号的关键下游效应物中，有一种被称为钙蛋白酶的蛋白质家族，它被Ca2+水平的变化直接激活。calpain是一种关键的蛋白酶，也是信号转导、迁移和细胞死亡的主要效应因子，但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和Ca2+如何影响细胞生理。我的这项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-利用生物信息学整合Ca2+与钙蛋白酶水解的作用。3-验证calpain切割的蛋白质，以更好地了解其生理作用。系统生物学为全新的探索途径创造了潜力，并在迭代周期中促进了不断的创新。蛋白水解和Ca2+信号是生物学的基本过程，但对它们的相互依赖性知之甚少。我的研究项目集中在使用尖端的蛋白质组学和生物信息学技术来研究Ca2+激活过程中调节的信号细胞网络，并解决这些关键的知识空白。该建议旨在表征Ca2+激活过程中切割的蛋白质，并补充我们目前对免疫细胞信号转导的了解。

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.