Expression, function and regulation of renal claudins

肾密蛋白的表达、功能和调节

• 批准号: 7847406
• 负责人: Alan S Yu
• 金额: $ 24.38万
• 依托单位: UNIVERSITY OF SOUTHERN CALIFORNIA
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-09-24 至 2011-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7847406
• 关键词: Affinity; Binding Sites; Cations; Cells; Charge; Coupled; Custom; Cysteine; Data; Development; Diabetes Insipidus; Disease; Drug Delivery Systems; Electrolytes; Electrostatics; Environment; Epithelium; Equipment; Excretory function; Extracellular Domain; Extracellular Space; Figs - dietary; Funding; Goals; Hypertension; Integral Membrane Protein; Ions; Kidney; Laboratories; Lateral; Lead; Location; Maps; Measurement; Membrane; Modeling; Monovalent Cations; Movement; Mutagenesis; Pathway interactions; Permeability; Play; Principal Investigator; Property; Regulation; Research; Role; Side; Site; Sodium Chloride; Structure; Structure-Activity Relationship; Testing; Tight Junctions; Water; Work; base; graduate student; mutant; novel; overexpression; parent grant; public health relevance; salt sensitive; urinary; wasting; water flow

DESCRIPTION (provided by applicant): The long-term goal of the parent grant is to determine the structural and functional determinants of paracellular permeation in epithelia. We previously developed a model using inducible overexpression of claudin-2 in MDCK I cells to study the properties of the claudin-2 cation pore. The parent grant had 3 specific aims, focused on characterizing the functional properties of claudin-2, identifying the basis of its cation selectivity, and mapping the structure of the pore. The impetus for this revision application is the recent discovery by our collaborator, Michael Fromm, that claudin-2 also substantially increases transepithelial water permeability. The current application seeks to expand the scope of the parent grant by adding a 4th specific aim, to characterize the mechanism of claudin-2-induced water permeability. This will generate new information that is itself entirely novel and physiologically significant, but will also add to the data generated by the other three specific aims to enhance our overall understanding of the structure of the pore. Furthermore, mutants generated in the original aims that alter ion selectivity and conductance can be exploited to probe the underlying mechanism of water permeability in this new aim. Thus, there will be substantial conceptual and experimental synergy between the proposed new aim and the original aims of the parent grant. This revision will accelerate the tempo of scientific research funded by the parent grant. It is expected to stimulate the economy by funding the hiring of a new graduate student, and procuring needed equipment for this project, including custom Ussing chambers for water flow measurements. PUBLIC HEALTH RELEVANCE: Principal Investigator: YU, Alan S. L. NARRATIVE This project seeks to understand the structure-function relationship of claudin-based pores and the mechanism for salt and water permeation. This may eventually lead to the identification of diseases that are due to abnormal paracellular permeability, which might include forms of salt-sensitive hypertension, salt-wasting, or diabetes insipidus. It may also lead to the development of drugs targeted to the claudin pore region that could modulate paracellular permeability and act as modulators of urinary electrolyte excretion.

描述（由申请人提供）：母基金的长期目标是确定上皮细胞中细胞旁渗透的结构和功能决定因素。我们先前开发了一种使用MDCK I细胞中紧密连接蛋白-2的诱导性过表达的模型来研究紧密连接蛋白-2阳离子孔的性质。母基金有3个具体目标，重点是表征claudin-2的功能特性，确定其阳离子选择性的基础，并绘制孔的结构。这一修订申请的动力是我们的合作者Michael Fromm最近发现的，即claudin-2也大大增加了跨上皮水渗透性。本申请试图通过增加第四个具体目标来扩大母授权的范围，以表征密蛋白-2诱导的水渗透性的机制。这将产生新的信息，这些信息本身是全新的，具有生理学意义，但也将增加其他三个特定目标产生的数据，以增强我们对孔结构的整体理解。此外，可以利用在原始目标中产生的改变离子选择性和电导的突变体来探索这个新目标中的水渗透性的潜在机制。因此，拟议的新目标与母补助金的原有目标在概念和实验方面将产生重大的协同作用。这一修订将加快由母基金资助的科学研究的克里思。预计将通过资助雇用一名新的研究生来刺激经济，并为该项目采购所需的设备，包括用于水流测量的定制尤辛室。 公共卫生相关性：主要研究者：YU，Alan S. L.叙述本项目旨在了解基于密蛋白的孔隙的结构-功能关系以及盐和水渗透的机制。这可能最终导致识别由于异常细胞旁通透性引起的疾病，其可能包括盐敏感性高血压、盐耗或尿崩症。这也可能导致开发针对密蛋白孔区域的药物，其可以调节细胞旁通透性并作为尿电解质排泄的调节剂。