Molecular Basis of Myocilin Function in the Human Eye

人眼肌纤蛋白功能的分子基础

• 批准号: 6518651
• 负责人: W Daniel Stamer
• 金额: $ 29.01万
• 依托单位: UNIVERSITY OF ARIZONA
• 依托单位国家: 美国
• 财政年份: 2001
• 资助国家: 美国
• 起止时间: 2001-07-02 至 2006-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6518651
• 关键词: cell adhesion molecules; cell cell interaction; glaucoma; green fluorescent proteins; human tissue; immunocytochemistry; immunoprecipitation; intercellular connection; intracellular transport; intraocular pressure; molecular biology; organ culture; protein structure function; protein transport; secretory protein; trabecular meshwork; transport proteins; vesicle /vacuole

DESCRIPTION (provided by applicant): The first molecular component involved in the pathogenesis of open-angle glaucoma has been identified. Mutations causing glaucoma in some are located to an open-angle glaucoma gene, GLC1A, on chromosome 1 that codes for a protein called myocilin. Myocilin is a novel protein of unknown function that is found in several eye tissues, including the trabecular meshwork, the likely site of pathology in open-angle glaucoma. On a subcellular level, myociin localizes to the vesicular compartment of trabecular meshwork cells, implicating vesicular traffic and/or secretion as affected pathways in open-angle glaucoma. Computer analyses of myocilin's primary sequence indicate significant homology to the SNARE [soluble N-ethylmaleimide sensitive fusion protein (NSF) attachment protein (SNAP) receptor] and Olfactomedin protein families, both of which localize to and function in the secretory pathway of cells. Moreover, preliminary data presented in this proposal indicate that 1) myocilin functions in both the formation and fusion of secretory-type vesicles in trabecular meshwork (HTM) cells and 2) myocilin-associated vesicles participate in the regulated assembly of nascent cell-cell junctions in two different epithelial cell models. Thus, the overall goal of this grant proposal is to determine the role of myocilin in the process of nascent cell-cell junction assembly in trabecular meshwork cells and the functional consequence of mutations in myocilin on this process. To this end, colocalization studies of myocilin with proteins known to participate in nascent cell-cell junctions will be performed in HTM cells. Second, the regulated targeting of myocilin to nascent cell-cell junctions in HTM cells will be studied. Finally, the functional consequence of mutations in myocilin (disease causing in humans) will be monitored using HTM cells as a model. These studies are designed to examine the molecular pathway in which myocilin functions in HTM cells. This knowledge will contribute to our understanding of glaucoma at the molecular level and provide new therapeutic targets for the treatment of those who are afflicted with glaucoma.

描述（申请人提供）：涉及的第一个分子成分 已经确定了开角式青光眼的发病机理。引起的突变 有些青光眼在某些人位于开放角度的青光眼基因GLC1A上 染色体1代码为一种称为肌动蛋白的蛋白质。 Myocilin是一本小说 在几种眼组织中发现的未知功能的蛋白质，包括 小梁网卫星，开放角度青光眼的病理可能位点。在 亚细胞水平，肌环素定位于小梁的囊泡区室 网格工作单元，牵涉到影响的囊泡流量和/或分泌 开角青光眼的途径。肌动蛋白主要的计算机分析 序列表明与SNARE [可溶性N-乙基马来酰亚胺 敏感融合蛋白（NSF）附着蛋白（SNAP）受体]和 嗅觉素蛋白家族都将其定位在 细胞的分泌途径。此外，在此提供的初步数据 提案表明1）肌动蛋白在形成和融合中起作用 小梁网（HTM）细胞中的分泌型囊泡和2） 与肌动蛋白相关的囊泡参与新生的调节组装 在两个不同的上皮细胞模型中的细胞细胞连接。因此，整体 该赠款提案的目标是确定肌动蛋白在此过程中的作用 小梁网细胞中的新生细胞 - 细胞连接组件和 在此过程中，肌动蛋白突变的功能后果。为此， 肌动蛋白与已知参与的蛋白质的共定位研究 新生的细胞 - 细胞连接将在HTM细胞中进行。第二， 在HTM细胞中，肌动蛋白对新生细胞连接的调节靶向 将被研究。最后，肌动蛋白突变的功能后果 （人类引起的疾病）将使用HTM细胞作为模型来监测。 这些研究旨在检查肌动蛋白的分子途径 在HTM细胞中的功能。这些知识将有助于我们对 分子水平的青光眼，并为 治疗患有青光眼的人。