BIOCHEMISTRY OF THE NEURONAL CYTOSKELETON

这项建议的总体目标是了解结构和 神经元细胞骨架的功能。建议的研究是设计的 让我们对神经元细胞骨架的功能有一个基本的了解 在正常的中枢神经系统。为此，我们将使用各种各样的 研究表达、组装、磷酸化和 一些神经元细胞骨架蛋白的相互作用。最终，我们 希望这些信息将帮助我们了解神经元为什么以及如何 细胞骨架会受到某些神经系统疾病的干扰，例如 阿尔茨海默病和肌萎缩侧索硬化症以及暴露于 神经毒素，如铝和丙烯酰胺。 这项建议的具体目的是1)研究相互作用和 三种神经丝蛋白在纯化的酶制剂中的组装 溴化氰衍生的每个神经丝三联体分子的片段 为了确定这三种蛋白的磷酸化在细胞周期中的作用 神经丝组装；2)鉴定和分离激酶和/或 神经细丝磷酸化的磷酸酶 蛋白质；3)继续我们对神经丝相互作用的研究 和微管，以确定哪些微管相关蛋白 (地图)参与了这些互动；4)研究 细胞骨架相关的ATPase，Beta-interexin在 细胞骨架介导的神经元和培养细胞的胞浆转运 5)获得高分子量神经细丝的cDNAs探针 蛋白质、核因子-M和核因子-H，将用于研究其表达和 对这些蛋白质的功能及各种修饰后的组装进行研究 蛋白质。这些克隆将被放入表达载体中，并将其 将在细胞中研究分布，细胞通常不包含 神经丝。这些研究将使我们能够确定哪些结构 它们在细胞中形成，它们的功能可能是什么。

The overall goal of this proposal is to understand the structure and function of the neuronal cytoskeleton. The proposed studies are designed to give us a basic knowledge of the function of the neuronal cytoskeleton in the normal CNS. Towards this end, we will use a wide variety of techniques to study the expression, assembly, phosphorylation and interactions of a number of neuronal cytoskeletal proteins. Ultimately, we hope that this information will help us understand why and how the neuronal cytoskeleton is perturbed by certain neurological diseases, such as Alzheimer's disease and amyotropic lateral sclerosis and by exposure to neurotoxins, such as aluminum and acrylamide. The specific aims of this proposal are 1) to study the interactions and assembly of the three neurofilament proteins using purified enzymatic and cyanogen bromide derived fragments of each neurofilament triplet molecule to determine the role of phosphorylation of these three proteins in the neurofilament assembly; 2) to identify and isolate the kinase and/or phosphatase responsible for the phosphorylation of the neurofilament proteins; 3) to continue our studies on the interactions of neurofilaments and microtubules, in order to define which microtubule associated proteins (MAPs) are involved in these interactions; 4) to study the importance of the cytoskeletal associated ATPase, Beta-internexin in cytoskeleton-mediated cytoplasmic transport in neurons and cultured cells and 5) to obtain cDNA probes for the high molecular weight neurofilament proteins, NF-M and NF-H, which will be used to study the expression and function of these proteins and to study the assembly of various modified proteins. The cDNA clones will be placed in expression vectors and their distribution will be studied in cells, which do not normally contain neurofilaments. These studies will allow us to determine what structures they form in the cell and what their function might be.