MOLECULAR FUNCTION OF SYNUCLEIN

描述（来自申请人的摘要）：该项目的长期目标 是定义突触核蛋白的正常和病理作用， 特别是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，因为一个明显的（但知之甚少）作用， 神经退行性疾病和正常的突触可塑性和学习。的 下一个项目期的第一个目标是完成一项正式的生物物理 人α-突触核蛋白（halphaS）的表征，重点是其 与膜脂相互作用并自缔合。这一目标将在 与巴塞尔大学的Seelig博士合作。第二个目标是 比较突变的突触核蛋白的性质与发展， 帕金森病，以及非人类（金丝雀）α-突触核蛋白，以获得 了解哪些特征是特别保守的，哪些可能更重要， 可能与病理学有关。第三个目标是使用定点诱变， 构建具有改变的脂质结合的重组halphaS的突变形式 特性.这将有助于深入了解 序列，并将产生潜在的突变体构建体用于探测 突触核蛋白的细胞功能第四个目标是绘制 突触核蛋白被蛋白质酪蛋白激酶II磷酸化，并研究 这种修饰对脂质结构和功能影响 结合和磷脂酶D2抑制。第五个目标将采用各种 在目的1-4中产生和表征的重组构建体，以探测 突触核蛋白对细胞功能发挥作用的机制。在 与马萨诸塞州总医院的海曼博士合作， 脂质结合对于细胞膜结合是必需假设将 的必要性，以及突触前和突触后的脂质结合的必要性。 还将测试末端定位和PLD 2抑制。总的来说， 这些实验验证了突触核蛋白的基本分子 功能与其保守的结构特征有关，这使其能够 与细胞内膜可逆结合。操纵这些 这种相互作用可以用于开发与年龄相关的 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.