Intracellular Transport: The Mannose Phosphate Receptor

细胞内运输：甘露糖磷酸受体

• 批准号: 7990866
• 负责人: Suzanne R Pfeffer
• 金额: $ 9.88万
• 依托单位: STANFORD UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-12-10 至 2011-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7990866
• 关键词: ATP Hydrolysis; ATP phosphohydrolase; Area; Back; Binding; Biochemical; Biological; Blood Circulation; Catalytic Domain; Cell physiology; Cells; Cholera Toxin; Cholesterol; Complex; Computer software; Confusion; Cultured Cells; Defect; Diabetes Mellitus; Disease; Early Endosome; Enzymes; Event; Genome; Goals; Golgi Apparatus; Guanosine Triphosphate Phosphohydrolases; Heart Diseases; Hela Cells; Human; IGF Type 2 Receptor; Immune system; Insulin; Intracellular Transport; Investigation; Lysosomes; Malignant Neoplasms; Mammalian Cell; Mannose; Mediating; Membrane Protein Traffic; Molecular; Monitor; Pathway interactions; Peripheral; Process; Protein Binding; Proteins; Recycling; Research; Role; Route; SNAP receptor; Small Interfering RNA; Sorting - Cell Movement; Surface; Testing; Transferrin Receptor; Transport Vesicles; Vesicle; Virus Diseases; Work; base; genome wide association study; genome-wide; inorganic phosphate; interest; killings; late endosome; nervous system disorder; novel; pathogen; protein function; public health relevance; receptor; receptor recycling; research study; syntaxin 6; trafficking; trans-Golgi Network

DESCRIPTION (provided by applicant): The long term goal of this research is to determine the molecular basis of membrane traffic in mammalian cells. The focus is on mannose 6-phosphate receptors (MPRs) that deliver newly synthesized lysosomal enzymes from the Golgi to pre-lysosomes, and then return to the Golgi to pick up more cargo. Several recently discovered proteins are needed for MPR transport from late endosomes to the trans Golgi network: a cargo selection protein that recognizes the MPRs in late endosomes (TIP47), a pathway- specific SNARE complex for fusion of MPR-vesicles at the TGN, and two proteins that function in vesicle tethering at the Golgi (GCC185 and RhoBTB3). The goals of this application are (1) to define precisely, the distinct routes taken by cargoes that are transported from early endosomes back to the Golgi, with focus on MPRs in comparison with cholera toxin; (2) to carry out a genome-wide, automated siRNA screen for proteins needed for MPR recycling. The screen will make use of the fact that depletion of proteins needed for MPR recycling leads to dispersal of MPRs into peripherally localized cellular compartments. Computer software can detect this dispersal, permitting automated analysis of the effects of 22,000 siRNAs transfected into cultured cells. (3) Also proposed are experiments to further characterize two novel Rab9 effectors that are important for this trafficking pathway: RhoBTB3 and RUTBC1. In summary, these experiments open up entirely new areas of investigation in the area of MPR trafficking and will provide fundamental information regarding the mechanisms of receptor trafficking in human cells. The work has broad application to our understanding of a number of disease states including diabetes, cancer, heart disease and neurological disorders. PUBLIC HEALTH RELEVANCE: Membrane traffic is essential for our ability to both secrete and respond to insulin, to clear cholesterol from the bloodstream, and for cells of the immune system to kill pathogens. Defects in membrane traffic underlie a number of disease states and virus infection depends upon this process. By understanding the molecular events responsible for membrane traffic, we will be better able to intervene in a variety of disease states.

描述（由申请人提供）：这项研究的长期目标是确定哺乳动物细胞中膜流量的分子基础。重点是将甘露糖6-磷酸受体（MPRS）从高尔基体传递新合成的溶酶体酶，然后返回高尔基体以捡起更多的货物。 MPR从晚期内体到Trans Golgi网络需要几种最近发现的蛋白质：一种识别晚期内体内MPR的货物选择蛋白（TIP47），这是一种途径 - 特定于TGN的MPR-Vesicles的途径 - 特异性的SNARE SNARE复合物，在TGN上融合了MPR-vess，并且在Golgi（Golgi）（GCC185）中起作用两种蛋白质的蛋白质。该应用的目标是（1）确切地定义，从早期内体转移到高尔基体的货物所采用的不同路线，与霍乱毒素相比，重点是MPRS； （2）进行全基因组，自动化的siRNA屏幕，以用于MPR回收所需的蛋白质。屏幕将利用以下事实：MPR回收所需的蛋白质的耗竭会导致MPRS分散到外围局部局部的细胞室中。计算机软件可以检测到这种扩散，从而可以自动分析转染到培养细胞中的22,000个siRNA的影响。 （3）还提出了实验，以进一步表征两个新型的RAB9效应子，这些效应因素对于该运输途径很重要：Rhobtb3和Rutbc1。总之，这些实验在MPR贩运领域开辟了全新的调查领域，并将提供有关人类细胞受体运输机制的基本信息。这项工作广泛地适用于我们对许多疾病状态（包括糖尿病，癌症，心脏病和神经系统疾病）的理解。公共卫生相关性：膜流量对于我们对胰岛素分泌和反应，从血液中清除胆固醇以及免疫系统细胞杀死病原体的能力至关重要。膜流量的缺陷是许多疾病状态和病毒感染的基础，取决于这一过程。通过了解负责膜流量的分子事件，我们将能够更好地干预各种疾病状态。