TOPOLOGY AND SUBCELLULAR LOBALIZATION OF PRESENILIN-1

描述(摘自申请者的摘要)：基因突变 编码早老素-1(PS1)蛋白在大多数 早发性家族性阿尔茨海默病(FAD)。因此，一个中心焦点是 阿尔茨海默病的研究是为了解决PS1在正常人群中的生理作用 和PS1相关的时尚大脑。最近的研究表明，PS1是一种跨膜蛋白 限制在细胞内膜隔间的蛋白质，在那里它经历 内切蛋白裂解成N-末端和C-末端片段。校长 本研究的目的是利用单抗的特异性。 识别PS1的N-末端和C-末端结构域及其融合的受体表位 PS1的连续截断，以解决PS1生物学的两个基本方面。 首先，表位保护试验和互补免疫金-EM研究 将用于确定野生型PS1的跨膜拓扑。 其次，将使用免疫金-EM来定义准确的 加工后PS1衍生物的超微结构定位。加在一起， 这些研究对于理解亚细胞间的相互作用至关重要。 PS1与其他分子沿着PS1的功能通路和沿 蛋白质本身的结构。我们还将研究以下方面的影响 选择野生型拓扑和天然N-端和C-端上的PS1突变 片段分布用于探索突变型PS1的潜在来源 故障。

DESCRIPTION (Adapted from applicant's abstract): Mutations in the gene encoding presenilin-1 (PS1) protein are causative in the majority of early-onset familial Alzheimer's disease (FAD). Thus, a central focus of Alzheimer's research is to resolve the physiological role of PS1 in normal and PS1-linked FAD brain. Recent studies show that PS1 is a transmembrane protein confined to intracellular membrane compartments, where it undergoes endoproteolytic cleavage into N- and C-terminal fragments. The principal goal of this research is to utilize the specificity of monoclonal antibodies recognizing N- and C-terminal domains of PS1 and a receptor epitope fused to serial truncations of PS1 to address two fundamental aspects of PS1 biology. First, an epitope protection assay and complementary immunogold-EM study will be adapted to determine the transmembrane topology of wild type PS1. Secondly, immunogold-EM will be employed to define the precise ultrastructural localization of processed PS1 derivatives. Taken together, these studies will be critical to understanding the subcellular interactions of PS1 with other molecules along the functional pathways of PS1 and along the structure of the protein itself. We will also examine the effects of select PS1 mutations on wild type topology and native N- and C-terminal fragment distribution to explore potential sources of mutant PS1 malfunction.