Liver Cell Membrane Protein-Expression and Function

总体描述(由申请人提供)： 这是一个计划项目，旨在继续研究特定肝细胞膜蛋白的功能和运输。提出了四个项目，涉及代表6个学术部门的15名教职调查人员。这些项目分享想法、技术和经常见面的调查人员，包括正式和非正式的。拟议的研究代表了对肝脏病理生物学中几个基本问题的多学科方法，包括膜蛋白在特定细胞骨架元件上的运输机制(项目1)，连接蛋白的表达和运输的调节(项目2)，内吞突变的运输步骤的确定(项目3)，以及运输到自噬空泡和溶酶体的机制(项目4)。项目4是新的，因为与现有的三个项目发展了强有力的合作，所以被添加到计划中。在过去五年中取得的主要成就包括：(项目1)含有单纯疱疹病毒的早期和晚期内吞小泡以及吐出小泡的基于微管的运动的体外重建，并确定了小泡相关的马达和调节因子；(项目2)鉴定和表征了连接蛋白32的蛋白质结合伙伴，它可能调节其进出质膜；(项目3)发现潜在的分选热休克蛋白异源复合体与磷酸化的去唾液酸糖蛋白受体胞浆结构域的关联是由新的酪蛋白激酶2α亚单位CK2(“”)介导的。该计划还包括三个核心设施。行政和支助事务核心(核心A)提供秘书、簿记和通用设备职能。分子细胞学核心(核心B)提供最先进的显微镜和成像服务。根据四个项目对这项技术的大量使用，增加了一个新的蛋白质组学核心(核心C)。所有四个项目都涉及肝细胞膜生物学和病理生物学的相互关联的领域，每个拟议的项目都代表了独立研究人员之间的合作努力。项目内部和项目之间的合作导致个人的专业知识扩展到肝脏病理生物学的重要领域，否则无法有效地进行研究。跟踪记录表明，参与调查的人员有能力在该计划的框架内协同工作和互动，以促进思想、方法和资源的共享。最终，本计划中进行的研究将带来基本的见解，这些见解对于理解和治疗或预防各种后天和遗传性肝脏疾病应该是重要的。

DESCRIPTION, OVERALL (provided by applicant): This is a Program Project to continue investigation of the functions and trafficking of specific liver cell membrane proteins. Four projects involving 15 faculty investigators representing 6 academic departments are proposed. These projects share ideas, techniques, and investigators who meet often, both formally and informally. The proposed studies represent multidisciplinary approaches to several fundamental questions in liver pathobiology including mechanisms of trafficking of membrane proteins on specific cytoskeletal elements (Project 1), the regulation of expression and trafficking of connexins (Project 2), the identification of trafficking steps in endocytosis mutants (Project 3), and mechanisms of trafficking to autophagic vacuoles and lysosomes (Project 4). Project 4 is new and was added to the Program because of strong collaborations that developed with the existing three Projects. Major accomplishments over the past 5 year period include: (Project 1) reconstitution in vitro of the microtubule-based motility of early and late endocytic vesicles as well as exocytosing vesicles that contain Herpes simplex virus and identification of vesicle-associated motors and regulatory factors; (Project 2) identification and characterization of protein binding partners for connexin 32 that may regulate its trafficking to and from the plasma membrane; (Project 3) discovery that the association of a potential sorting heat shock protein heterocomplex with the phosphorylated asialoglycoprotein receptor cytoplasmic domain is mediated by a novel casein kinase 2 alpha subunit, CK2(". This Program also includes three Core facilities. The Administrative and Supporting Services Core (Core A) provides secretarial, bookkeeping, and common equipment functions. The Molecular Cytology Core (Core B) provides state-of-the-art microscopy and imaging services. A new Proteomics Core (Core C) has been added based upon the substantial use of this technology by each of the four Projects. All four Projects are concerned with interrelated areas of hepatocyte membrane biology and pathobiology, and each proposed project represents collaborative efforts between independent investigators. Collaborations within and between projects lead to extensions of an individual's expertise into important areas of liver pathobiology that could not otherwise be effectively investigated. The track record demonstrates the ability of the investigators involved to work together and interact synergistically within the framework of this Program to promote the sharing of ideas, methodology, and resources. Ultimately, the studies performed in this Program will lead to fundamental insights that should be important for understanding and treating or preventing various acquired and inherited disorders of the liver.