Calcium-Dependent Protein Phosphorylation in the Green Alga Mougeotia

本研究探讨了钙的作用机制， 植物细胞 钙是一种非常重要的调节分子， 所有的真核生物。 它被认为是细胞内的一个重要的细胞因子。 将生理反应与外部 刺激。 钙结合蛋白已经被广泛地 在动物身上研究。 在这个项目中， 将在植物中研究钙的作用。 的目标 研究是为了促进我们对钙如何介导 环境残株对植物细胞的影响。 选择绿色转板藻作为实验材料 因为它是一个有吸引力的钙生物模型- 植物的依赖性调节 该方法将涉及 分子靶点的纯化和表征 转板藻中的钙：钙刺激的蛋白质和酶。 重点将放在最近发现的钙刺激 转板藻属蛋白激酶。 纯化和 将进行蛋白激酶的表征。 特别强调的是分子的阐明， 通过识别钙结合的钙调节的基础 调节活性的效应分子。 此外，为了 确定这种激酶在转板藻中的作用，在体外和在 体内磷酸化测定将用于鉴定 蛋白激酶的底物。 磷酸化的影响 将评估这些底物的体外活性。 体内底物磷酸化水平的变化， 触发或抑制钙依赖性反应的条件 也将被确定。

This research investigates the mechanisms of calcium action in plant cells. Calcium is a very important regulatory molecule in all eukaryotes. It is thought to play a role as an intracellular messenger that couples physiological responses to external stimuli. The calcium binding proteins have been extensively studied in animals. In this project, the potential targets of calcium action will be investigated in plants. The goal of the research is to advance our understanding of how calcium mediates the effects of environmental stumuli on plant cells. The green alga Mougeotia has been selected as the experimental organism because it is an attractive biological model of calcium- dependent regulation in plants. The approach will involve purification and characterization of the molecular targets of calcium in Mougeotia: calcium-stimulated proteins and enzymes. The focus will be on a recently discovered calcium stimulated protein kinase of Mougeotia. The purification and characterization of the protein kinase will be carried out. Particular emphasis will be on the elucidation of the molecular basis of calcium regulation by identifying the calcium-binding effector molecules that regulate the activity. Also, in order to determine the role of this kinase in Mougeotia, in vitro and in vivo phosphorylation assays will be used to identify the substrates of the protein kinase. The effects of phosphorylation on the in vitro activities of these substrates will be assessed. The changes in in vivo substrate phosphorylation levels under conditions that trigger or suppress calcium-dependent responses also will be determined.//