INTRACELLULAR TRANSPORT--MANNOSE-PHOSPHATE RECEPTOR

细胞内转运--甘露糖磷酸受体

• 批准号: 3236198
• 负责人: Suzanne R Pfeffer
• 金额: $ 20.64万
• 依托单位: STANFORD UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 1986
• 资助国家: 美国
• 起止时间: 1986-07-01 至 1994-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3236198
• 关键词: Golgi apparatus; affinity chromatography; binding proteins; cell membrane; clathrin; glycoproteins; hamsters; intracellular membranes; intracellular transport; laboratory rabbit; lysosomes; mannose; membrane proteins; protein purification; protein transport; radioimmunoassay; vesicle /vacuole

During intracellular transport, proteins destined for the plasma membrane, secretory vesicles, and lysosomes must be sorted from one another within the Golgi complex, and sent to their appropriate addresses. The long term goal of this research is to elucidate the molecular mechanisms by which proteins are targeted to specific and distinct intracellular destinations. The targeting of proteins to lysosomes is presently the best understood sorting process. Soluble lysosomal enzymes acquire a mannose 6-phosphate (man6P) tag; intracellular receptors recognize man6P-containing proteins, and somehow facilitate their transport to lysosomes. The specific aims of this proposal are: 1. To study protein sorting in the trans Golgi network (TGN), clathrin coated vesicles carrying man6P receptors will be isolated by immunoadsorption. Their content of a lysosomal membrane glycoprotein will then be analyzed to determine if these two proteins are transported together from the TGN to pre-lysosomes. 2. To study protein sorting in pre-lysosomes, the potential recycling of a lysosomal membrane glycoprotein from pre-lysosomes to the TGN will be investigated. The ability of a soluble lysosomal enzyme to be transported to the TGN will also be examined. 3. To study the molecular basic of vesicular transport from pre-lysosomes to the TGN, an analysis of the cytosolic factors that facilitate this process will be initiated using a cell-free system developed in this laboratory, Purification of an N-ethylmaleimide-sensitive factor will attempted. Experiments will be carried out to examine the potential role of clathrin and a transport factor termed "NSF" in this transport process. Finally, an anti-peptide antiserum will be generated to identify a family of GTP binding proteins that facilitates intracellular transport. It is hoped that these cell biological and biochemical experiments will further our understanding of the molecular mechanisms by which proteins are transported between membrane-bound compartments within the cytoplasm of eukaryotic cells.

在细胞内运输过程中，蛋白质注定要用于质膜， 分泌囊泡和溶酶体必须在内部彼此分类 高尔基体综合大楼，并发送到其适当的地址。长期 这项研究的目标是阐明分子机制 蛋白质针对特定和不同的细胞内目的地。 蛋白质靶向溶酶体目前是最好的理解 排序过程。可溶性溶酶体酶获得6-磷酸甘露糖 （man6p）标签；细胞内受体识别含MAN6P的蛋白质， 并以某种方式促进了他们运输到溶酶体。具体目的 该提议是：1。在反式高尔基网络中研究蛋白质排序 （TGN）将分离出携带MAN6P受体的网状蛋白涂层囊泡 通过免疫吸附。它们的溶酶体膜糖蛋白的含量 然后将分析以确定是否运输这两种蛋白质 从TGN到前溶酶体在一起。 2。在研究前溶酶体中的蛋白质分类，A的潜在回收 溶酶体膜糖蛋白从前溶质体到TGN将是 调查。可溶性溶酶体酶运输的能力 也将检查到TGN。 3。研究来自前溶质体的囊泡转运的分子基础 对于TGN，对促进这一点的胞质因子的分析 过程将使用此中开发的无单元系统启动 实验室，纯化N-乙基马来酰亚胺敏感因素将 尝试。实验将进行检查以检查 在此运输过程中，网状蛋白和一个被称为“ NSF”的传输因子。 最后，将生成抗肽抗血清以识别一个家庭 促进细胞内转运的GTP结合蛋白。 希望这些细胞生物学和生化实验能够 进一步我们对蛋白质的分子机制的理解 在细胞质内的膜结合室之间运输 真核细胞。