MAMMALIAN COG4

哺乳动物COG4

• 批准号: 7957255
• 负责人: FREDERICK M HUGHSON
• 金额: $ 0.79万
• 依托单位: BROOKHAVEN SCIENCE ASSOC-BROOKHAVEN LAB
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-07-01 至 2010-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7957255
• 关键词: Affect; Architecture; Area; Cell Surface Proteins; Complex; Computer Retrieval of Information on Scientific Projects Database; Crystallography; Data; Data Collection; Defect; Funding; Goals; Golgi Apparatus; Grant; Image; Institution; L-Selenomethionine; Laboratories; Light; Membrane; Molecular; Pathway interactions; Phase; Play; Proteins; Publishing; Research; Research Personnel; Resources; Role; Science; Sorting - Cell Movement; Source; Structure; Synchrotrons; United States National Institutes of Health; Vesicle; Work; Writing; detector; glycosylation; human disease; meetings; trafficking

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. The Golgi apparatus plays a key role in protein sorting and glycoslyation within the eukaryotic secretory pathway. Defects in vesicular trafficking within the Golgi affect both its structure and function. As a consequence, such defects can have pleiotropic effects on the glycosylation and stability of cell surface proteins, leading to human disease. This proposal arises from work funded by the NIH (RO1 GM071574: "Structural Analysis of Golgi Tethering Proteins"). Our efforts in this area, generally speaking, are focused on two large complexes, the Conserved Oligomeric Golgi (COG) complex and the Dsl1 complex. Both are essential for intracellular trafficking and are thought to act in the initial tethering of intracellular trafficking vesicles to their membrane targets. Nonetheless, despite intensive efforts, little is known about the molecular mechanism by which these complexes function in vesicle tethering. Our long-term goal is to determine crystal structures of subunits and subassemblies of the COG (8 subunits) and Dsl1 (4 subunits) complexes. We have done extensive preliminary work, especially with COG, in characterizing the overall architecture (i.e. subunit connectivity) of the complex. There are, however, no published structures of any portion of either complex. We have recently obtained promising crystals of subunits of each complex, and propose to determine their structures at NSLS using MAD phasing. Here, we write to request X-25 or X-29 beamtime to determine the structure of a fragment of mammalian Cog4. We call this 265-residue fragment Cog4B. Our current Cog4B crystals diffract x-rays well, to at least 3.0 angstroms using our laboratory's R-Axis IV image plate detector. Therefore, current efforts are focused on obtaining SeMet-substituted crystals for MAD data collection. Since the total number of Met residues is low for a protein of this size (4 Met over 265 residues), high-quality synchrotron data is likely to be instrumental for MAD phasing. Given the importance of the science and the promise of the preliminary data, we feel it is vital to collect data as soon as possible.

该副本是利用众多研究子项目之一 由NIH/NCRR资助的中心赠款提供的资源。子弹和 调查员（PI）可能已经从其他NIH来源获得了主要资金， 因此可以在其他清晰的条目中代表。列出的机构是 对于中心，这不一定是调查员的机构。 高尔基体在真核分泌途径内的蛋白质分类和糖脂化中起关键作用。 高尔基体内囊泡运输的缺陷会影响其结构和功能。 因此，这种缺陷可以对细胞表面蛋白的糖基化和稳定性产生多效性影响，从而导致人类疾病。 该提案是由NIH资助的工作（RO1 GM071574：“高尔基链接蛋白的结构分析”）产生的。 一般而言，我们在这一领域的努力集中在两个大型复合物上，即保守的低聚高尔基（COG）复合物和DSL1复合物。 两者都是细胞内贩运必不可少的，并且被认为是将细胞内贩运囊泡最初束缚到其膜靶标的作用。 尽管如此，尽管进行了大量努力，但对这些复合物在囊泡束缚中起作用的分子机制知之甚少。 我们的长期目标是确定COG（8个亚基）和DSL1（4个亚基）配合物的亚基和亚组件的晶体结构。 我们已经完成了广泛的初步工作，尤其是在COG的情况下，在表征该综合体的整体结构（即子单位连接性）方面。 但是，没有任何一个复合物的任何部分发表的结构。 最近，我们获得了每个复合物的亚基的有希望的晶体，并建议使用MAD逐步确定其在NSL的结构。 在这里，我们写信以请求X-25或X-29 Beamtime，以确定哺乳动物COG4片段的结构。 我们称这个265个残留的片段COG4B。 我们当前的COG4B晶体衍射X射线良好，使用实验室的R轴IV图像板检测器至少3.0埃。 因此，当前的工作集中在获得疯狂数据收集的Semet取代晶体。 由于这种大小的蛋白质的MET残基总数很少（4个MET超过265个残基），因此高质量的同步加速器数据可能对疯狂的相位有用。 鉴于科学的重要性和初步数据的希望，我们认为尽快收集数据至关重要。