ANNEXIN PHOSPHORYLATION: CAUSES AND CONSEQUENCES

描述(申请人摘要)：膜联蛋白是钙依赖的， 磷脂结合蛋白是运动的主要外周蛋白 对细胞内钙信号事件作出反应的膜上和膜外。 有证据表明，它们调节细胞膜上的许多酶活性，如 作为蛋白激酶和磷脂酶，它们充当受体和对接位置 对于膜上的其他蛋白质，它们可能介导膜-膜 胞吐作用、胞吞作用和细胞器转运的相互作用。一个 一些膜联蛋白被磷酸化，以响应细胞刺激 分泌物、生长因子和致癌因子。这一项目旨在 通过识别膜联蛋白的最重要的功能 它们被磷酸化的生理和病理条件， 所涉及的蛋白激酶，以及磷酸化的后果 关于调节细胞活动的条款。膜联蛋白VI的结构 一种模拟磷酸化的突变将通过X射线确定 结晶学。天然和突变型膜联蛋白VI在脂质上的构象 单分子膜将由二维晶体的电子显微镜来确定。 与膜联蛋白上的磷酸化位点对接的蛋白质将是 特色化的。磷酸化对细胞定位和运动的影响 将确定细胞中的膜联蛋白，包括膜联蛋白的运动 进入细胞的细胞核。这些关于组织的基础研究 膜及其相关蛋白质应该有助于更好地理解 细胞信号事件是荷尔蒙和 神经递质与细胞生长调控 病理情况。

DESCRIPTION (Applicant's abstract): The annexins are calcium-dependent, phospholipid-binding proteins that are the major peripheral proteins that move on and off membranes in response to calcium signalling events in cells. Evidence suggests they regulate a number of enzyme activities on membranes such as protein kinases and phospholipases, they act as receptors and docking sites for other proteins on the membrane, and they may mediate membrane-membrane interactions underlying exocytosis, endocytosis, and organelle tra.fficking. A number of annexins are phosphorylated in response to cell stimulation by secretogogues, growth factors, and oncogenic kinases. This project seeks to understand the most important ftinctions of annexins by identifying the physiological and pathological conditions under which they are phosphorylated, the protein kinases involved, and the consequences of the phosphorylation in terms of the regulation of cellular activities. The structure of annexin VI with a mutation that mimics phosphorylation will be determined by X-ray crystallography. The conformation of native and mutant annexin VI on lipid monolayers will be determined by electron microscopy of 2 dimensional crystals. Proteins that dock onto the phosphorylation sites on annexins will be characterized. The influence of phosphorylation on the locations and movements of annexins in cells will be determined, including the movements of annexins into the nucleus of the cell. These fundamental studies on the organization of membranes and their associated proteins should lead to a better understanding of cell signalling events that underlie the release of hormones and neurotransmitters and the regulation of cell growth under normal and pathological conditions.