NEUROFIBROUS PROTEINS IN NORMAL AND AGING BRAIN

正常和衰老大脑中的神经纤维蛋白

• 批准号: 3409168
• 负责人: FELICIA GASKIN
• 金额: $ 17.8万
• 依托单位: UNIVERSITY OF VIRGINIA
• 依托单位国家: 美国
• 财政年份: 1988
• 资助国家: 美国
• 起止时间: 1988-12-01 至 1994-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3409168
• 关键词: Alzheimer's disease; adenosinetriphosphatase; aging; brain metabolism; cell type; electron microscopy; enzyme mechanism; fibrous protein; guanosine diphosphate; high performance liquid chromatography; histochemistry /cytochemistry; insulin receptor; kinesin; laboratory mouse; laboratory rabbit; laboratory rat; microtubule associated protein; microtubules; monoclonal antibody; nuclear magnetic resonance spectroscopy; nucleoside diphosphate kinase; nucleotide analog; nucleotides; phosphorylation; polymerization; posttranslational modifications; protein biosynthesis; radiotracer; tubulin

The overall aim of this research is to understand the role of nucleotides, microtubule associated proteins (MAPS) and posttranslational modifications in the regulation of microtubule-mediated functions in cells. There is increasing evidence that co-factors, MAPs and posttranslational modifications of tubulin and MAPs show age-dependent changes and some of these changes may be important in Alzheimer's Disease. Specific Aims include: 1. To continue to probe microtubule assembly in vitro using thiophosphate analogues of GTP and ATP and highly purified tubulin free of contaminating nucleotide diphosphate kinase (NDPK) and ATPase activities to further our understanding on the mechanism of nucleotide binding and hydrolysis in tubulin assembly. In addition, the effects of tubulin-stimulated ATPase (MAP-lC), kinesin (another ATPase that reacts with tubulin) and NDPK will be probed in the microtubule assembly system with thiophosphate analogues of nucleotides. 2. To use tubulin tyrosine ligase or insulin receptor kinase to add phosphotyrosine or to phosphorylate tyrosine at the carboxy terminal end of alpha-tubulin. To analyze the potential role of phosphorylation and dephosphorylation of this critical tyrosine in tubulin assembly. 3. To further characterize the mechanism of action of tubulin and MAP-2 regulation on the phosphotyrosyl phosphatase activity of the low-molecular weight polycation-stimulated protein phosphatase. These studies should help to elucidate the role of nucleotides, microtubule associated enzymes and phosphorylation on the cytoskeleton and increase our understanding of microtubule functions. They should also contribute to our understanding of age-dependent alterations of microtubules in brain.

这项研究的总体目标是了解核苷酸的作用， 微管相关蛋白（MAPS）和翻译后修饰 在调节细胞中微管介导的功能方面。 有 越来越多的证据表明，辅因子，地图和翻译后 微管蛋白和MAP的修饰显示出年龄依赖性变化， 这些变化可能对阿尔茨海默病很重要。 具体目标 有：1.为了继续探索微管组装在体外使用 GTP和ATP的硫代磷酸类似物以及不含 污染的核苷酸二磷酸激酶（NDPK）和ATP酶活性 为了进一步了解核苷酸结合的机制， 微管蛋白组装中的水解。 此外， 微管蛋白刺激的ATP酶（MAP-1C）、驱动蛋白（另一种与微管蛋白反应的ATP酶）、微管蛋白刺激的ATP酶（MAP-1C）、微管蛋白刺激的ATP酶（另一种与微管蛋白反应的ATP酶）、微管蛋白刺激的ATP酶（MAP-1C）、微管蛋白刺激的ATP酶（另一种与微管蛋白反应的ATP酶）、微管蛋白刺激的ATP酶（MAP-1C）、微管蛋白刺激的ATP酶（另一种与微管蛋白反应的ATP酶）、微管蛋白刺激的ATP酶（另一种与微管蛋白反应的ATP酶）、微管蛋白刺激的ATP酶（另一种与微管蛋白反应的ATP酶）、微管蛋白刺激的ATP酶（另一种与微管蛋白反应的ATP酶）。 与微管蛋白）和NDPK将在微管组装系统中进行探测 与核苷酸的硫代磷酸酯类似物。2.利用微管蛋白酪氨酸 连接酶或胰岛素受体激酶添加磷酸酪氨酸或 在α-微管蛋白的羧基末端磷酸化酪氨酸。 到 分析磷酸化和去磷酸化的潜在作用， 微管蛋白装配中的关键酪氨酸。3.为了进一步表征 微管蛋白和MAP-2调节磷酸酪氨酸的作用机制 低分子量聚阳离子刺激的磷酸酶活性 蛋白磷酸酶 这些研究应有助于阐明 核苷酸，微管相关酶和磷酸化对 细胞骨架和增加我们对微管功能的理解。 他们 也有助于我们理解年龄依赖性的变化 大脑中的微管。