MOLECULAR FUNCTION OF SYNUCLEIN

描述(摘自申请者的摘要)：该项目的长期目标 是定义突触核蛋白的正常和病理作用，以及在 特别的α-突触核蛋白。α-突触核蛋白已经成为一个主要的焦点 调查是因为在这两个事件中都扮演了明显(但了解不多)的角色 神经退行性疾病和正常突触可塑性和学习。这个 下一个项目阶段的第一个目标是完成正式的生物物理 人α-突触核蛋白(HalphaS)的特性，重点是它的能力 与膜脂相互作用并自我结合。这一目标将被实现 与巴塞尔大学的Seelig博士合作。第二个目标是 比较突变型突触核蛋白与血管内皮细胞瘤发生的关系 帕金森病，以及一种非人类(金丝雀)α-突触核蛋白，以获得 洞察哪些功能被特别保留，哪些功能可能更多 很可能与病理学有关。第三个目标是使用定点突变来 构建脂结合改变的重组HalphaS突变体 属性。这将使我们能够深入了解 序列，并将生成可能对探测有用的突变结构 突触核蛋白的细胞功能。第四个目标是绘制出 突触核蛋白被酪蛋白激酶II磷酸化，并研究 这种修饰对脂质的结构和功能的影响 结合和磷脂酶D2抑制。第五个目标将使用各种不同的 在AIMS 1-4中产生和表征的重组载体，以探索 突触核蛋白对细胞功能发挥作用的机制。在……里面 与马萨诸塞州综合医院的海曼博士合作 假设脂质结合是细胞膜结合所必需的 接受正式测试，以及两种突触前的脂质结合的必要性 还将测试末端定位和PLD2抑制。总而言之， 这些实验验证了这样一个假设，即突触核蛋白的基本分子 功能与其保守的结构特征有关，这使得它能够 与细胞内膜可逆结合。对这些的操控 互动在开发与年龄相关的治疗方法方面可能会有用处 包括阿尔茨海默氏症和帕金森氏症在内的疾病。

DESCRIPTION (From the applicant's abstract): The long-term goal of this project is to define the normal and pathological roles of the synucleins, and in particular alpha-synuclein. Alpha-synuclein has emerged as a major focus of investigation because of an apparent (but poorly understood) role both in neurodegenerative disease and in normal synaptic plasticity and learning. The first aim for the next project period is to complete a formal biophysical characterization of human alpha-synuclein (halphaS), focusing on its ability to interact with membrane lipids and to self-associate. This aim will be conducted in collaboration with Dr. Seelig of the University of Basel. A second aim is to compare the properties of mutant synucleins linked to the development of Parkinson's disease, as well as a non-human (canary) alpha-synuclein, to gain insight into which features are specifically conserved and which may be more likely related to pathology. A third aim is to use site-directed mutagenesis to construct mutant forms of recombinant halphaS with altered lipid binding properties. This will provide insight into the functional organization of the sequence, and will generate mutant constructs potentially useful for probing synuclein's cellular functions. A fourth aim is to map the sites upon which synuclein is phosphorylated by the protein Casein Kinase II, and to investigate the structural and functional consequences of this modification on lipid binding and phospholipase D2 inhibition. A fifth aim will employ the various recombinant constructs produced and characterized in Aims 1-4, to probe the mechanisms by which synuclein can exert effects on cell function. In collaboration with Dr. Hyman of the Massachusetts General Hospital, the hypothesis that lipid binding is necessary for cell membrane association will be formally tested, and the necessity of lipid binding for both presynaptic terminal localization and PLD2 inhibition will also be tested. Collectively, these experiments test the hypothesis that synuclein's essential molecular function is related to its conserved structural features, which allow it to bind reversibly with intracellular membranes. Manipulation of these interactions could have uses in the development of therapies for age-related diseases including Alzheimer's and Parkinson's diseases.