Structure-Function of Type IV Collagen NC1 Domains

IV 型胶原 NC1 结构域的结构-功能

• 批准号: 6844677
• 负责人: MUNIRATHINAM SUNDARAMOORTHY
• 金额: $ 22.57万
• 依托单位: VANDERBILT UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2002
• 资助国家: 美国
• 起止时间: 2002-05-01 至 2007-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6844677
• 关键词: Goodpasture' s syndrome; X ray crystallography; angiogenesis inhibitors; antigens; autoantibody; basement membrane; chemical structure; collagen; computer simulation; crystallization; molecular dynamics; molecular site; protein isoforms; protein structure function; renal glomerulus; stoichiometry; structural biology; triple helix

DESCRIPTION (provided by applicant): The structure of glomerular basement membrane (GBM) is altered in three notable diseases that affect the kidneys: Goodpasture syndrome (anti-GBM nephritis), Alport syndrome (hereditary nephritis), and diabetes mellitus. At least two of them, Goodpasture and Alport syndromes, have been linked to type IV collagen, a major constituent of GBM. Type IV collagen is a family of six alpha chains, alpha1-alpha6, found in various basement membranes (BMs). Three alpha chains form a collagen triple helical protomer and each chain contains a noncollagenous (NC1) globular domain at the C-terminus. The chain stoichiometry of the protomer varies with the origin of the tissue, (alpha1)2.alpha2 being the ubiquitous form and alpha3.alpha4.alpha5 being specific to glomeruli and alveoli. Two triple helical protomers associate at the NC1 region to form tail-to-tail dimer. The NC1 domains are presumed to contain the structural determinants for both triple helix formation and hexamer assembly. The alpha3 NC1 domain harbors the epitope for autoantibodies in patients with Goodpasture syndrome. Mutations in the genes encoding any of the three alpha chains result in the loss of alpha3.alpha4.alpha5 network in the GBM, which is the cause of Alport syndrome. Recombinant alpha2, alpha3, and alpha6 NC1 domains show anti-angiogenic and anti-tumor properties. In this study, we propose the following specific aims to determine the structure-function relationships of NC1 domains: Aim 1: To determine the first crystal structure of a NC1 hexamer, ((alpha1)2.alpha2)2 Aim 2: To crystallize and determine the structure of GBM NC1 hexamer, (alpha3.alpha4.alpha5)2 Aim 3: To determine the structure of the Goodpasture antigen, alpha3 NC1 monomer Aim 4: To solve the structures of alpha2 and alpha6 monomers, the angiogenic inhibitors. The accomplishment of these aims requires the application of macromolecular crystallography, computational biology, protein chemistry and molecular biology.

描述（由申请人提供）：肾小球地下室的结构 在影响肾脏的三种著名疾病中，膜（GBM）发生了变化： Goodpasture综合征（抗GBM肾炎），Alport综合征（遗传 肾炎）和糖尿病。其中至少两个，Goodpasture和Alport 综合征已与IV型胶原蛋白（GBM的主要组成部分）联系起来。 IV型胶原蛋白是一个六个alpha链的家族，Alpha1-alpha6在 各种地下膜（BMS）。三个α链形成胶原蛋白三重 螺旋原型和每个链都包含一个非胶原（NC1）球状结构域 在C末端。毒剂的链化学计量法随着 组织的起源，（alpha1）2。α2是无处不在的形式和 alpha3.alpha4.Alpha5特定于肾小球和肺泡。两个三倍 在NC1区域辅助螺旋型原子物形成尾巴二聚体。这 假定NC1结构域包含两个三重的结构决定因素 螺旋形成和六聚体组件。 Alpha3 NC1域藏有表位 用于良好综合征患者的自身抗体。突变 编码三个α链中任何一个的基因导致丧失 GBM中的Alpha3.alpha4.alpha5网络，这是Alport综合征的原因。 重组α2，α3和α6NC1结构域显示抗血管生成和 抗肿瘤特性。在这项研究中，我们提出以下具体目的 确定NC1域的结构功能关系： 目标1：确定NC1六聚体的第一个晶体结构， （（alpha1）2.Alpha2）2 目标2：要结晶并确定GBM NC1六聚体的结构， （alpha3.alpha4.alpha5）2 目标3：确定固定抗原的结构，alpha3 nc1 单体 目标4：解决α2和α6单体的结构，即血管生成 抑制剂。 这些目标的完成需要应用大分子 晶体学，计算生物学，蛋白质化学和分子 生物学。