Structure and Function of the Exocyst Complex

外囊复合体的结构和功能

基本信息

批准号：
9298681
负责人：
Mary Munson
金额：
$ 50.74万
依托单位：
UNIV OF MASSACHUSETTS MED SCH WORCESTER
依托单位国家：
美国
项目类别：
财政年份：
2005
资助国家：
美国
起止时间：
2005-07-01 至 2018-09-20
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9298681
关键词：
Animal Model Architecture Binding Biochemical Biological Biological Process Biophysics Cell membrane Cells Cellular Morphology Cellular biology Chimeric Proteins Communication Complement Complex Crystallography Data Development Diabetes Mellitus Dissection Electron Microscopy Eukaryota Eukaryotic Cell Event Foundations Funding Genetic Growth Growth and Development function Hormones Human Imagery In Vitro Individual Investigation Knowledge Lead Malignant Neoplasms Maps Mass Spectrum Analysis Measures Mechanics Mediating Membrane Membrane Fusion Methods Modeling Molecular Molecular Conformation Molecular Models Molecular Structure Monomeric GTP-Binding Proteins Movement Negative Staining Neurons Organelles Pathogenesis Pathway interactions Plant Roots Process Proteins Publishing Quality Control Reagent Regulation Research Resolution Roentgen Rays Role SNAP receptor Saccharomyces cerevisiae Secretory Vesicles Site Specificity Spectrum Analysis Structure Techniques Technology Testing Time Vesicle Vesicle Transport Pathway Yeast Model System Yeasts biophysical techniques cell growth ciliopathy crosslink human disease improved in vivo insight man molecular modeling multidisciplinary mutant neurotransmission novel protein complex public health relevance rab GTP-Binding Proteins rho GTP-Binding Proteins single molecule three dimensional structure tool trafficking yeast genetics

项目摘要

DESCRIPTION (provided by applicant): Eukaryotic cells transport cargo between subcellular organelles, and to the plasma membrane for secretion, using small membrane-bound vesicles are carriers. The regulation of vesicular transport and membrane fusion processes are crucial for cellular morphology, growth, movement and secretion, including hormone release and neurotransmission. Many essential proteins are required for these processes, including the SNARE proteins and Sec1 that are involved in the membrane fusion process, the Rab and Rho GTPases, and an octameric tethering complex called the exocyst. Although the exocyst complex has been implicated in a number of different functions involved in recognition, tethering and quality control of SNARE assembly and fusion, none of these are well understood at the molecular level. We are using a multidisciplinary strategy of biochemical and biophysical techniques, combined with genetics and cell biological methods, in order to understand the molecular architecture and function of the exocyst complex. We study the exocyst proteins from the model organism Saccharomyces cerevisiae to take advantage of the wealth of genetic, cell biological and biochemical techniques available. Our studies aim to: 1) map the functional organization of the exocyst complex through biochemical studies in vitro and analyze mutants to test the function of the exocyst in vivo; (2) determine the 3D structure of the entire exocyst complex using electron microscopy, crystallography and molecular modeling; and (3) watch the exocyst tether vesicles at the single molecule level to analyze the requirements for tethering, and (4) dissect the role of the exocyst and Sec1 in SNARE complex assembly and membrane fusion. Because these proteins are conserved from yeast to human neurons, this research will advance our knowledge of how secretion and growth are regulated in all eukaryotic cells.

描述（由申请人提供）：真核细胞使用小膜结合的小囊泡在亚细胞器之间和质膜之间运输货物进行分泌。囊泡转运和膜融合过程的调节对于细胞形态，生长，运动和分泌，包括激素释放和神经传递至关重要。这些过程需要许多必需的蛋白质，包括膜融合过程中涉及的SNARE蛋白和SEC1，RAB和RHO GTPases以及一个称为外旋转的八接束缚络合物。尽管外囊复合物已与识别，绑扎和质量控制的许多不同功能有关，但在分子水平上都没有很好地理解这些功能。我们正在使用生物化学和生物物理技术的多学科策略，结合遗传学和细胞生物学方法，以了解胞外循环复合物的分子结构和功能。我们研究了酿酒酵母的模型生物体中的胞外蛋白，以利用可用的遗传，细胞生物学和生化技术的丰富。我们的研究目的是：1）通过体外的生化研究来绘制外囊肿复合物的功能组织，并分析突变体以测试体内外囊肿的功能；（2）使用电子显微镜，晶体学和分子建模来确定整个胞外复合物的3D结构；（3）观察单分子水平的外旋囊泡以分析绑扎的需求，（4）剖析外囊肿和SEC1在SNARE COMPLECS组装和膜融合中的作用。由于这些蛋白质是从酵母到人神经元保守的，因此这项研究将促进我们对所有真核细胞中分泌和生长如何受到调节的了解。