CRYSTAL STRUCTURE DETERMINATION OF COLLAGEN-RELATED STRUCTURES

胶原相关结构的晶体结构测定

• 批准号: 7721798
• 负责人: MUNIRATHINAM SUNDARAMOORTHY
• 金额: $ 0.02万
• 依托单位: STANFORD UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2008
• 资助国家: 美国
• 起止时间: 2008-03-01 至 2009-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7721798
• 关键词: Antibodies; Autoimmune Diseases; Basement membrane; C-terminal; Collagen; Collagen Type IV; Computer Retrieval of Information on Scientific Projects Database; Data; Development; Extracellular Matrix; Funding; Goodpasture Syndrome; Grant; Hereditary nephritis; Housing; Institution; Kidney; Kidney Diseases; Link; Mammals; Methods; Modeling; Molecular; Molecular Structure; Pathogenesis; Play; Protomer; Research; Research Personnel; Resources; Role; Source; Structural Protein; Structure; Tissues; United States National Institutes of Health; base; interest; monomer

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. Type IV collagen is a major structural protein component of basement membrane of all metazoan. It contains a large C-terminal globular domain (NC1 domain) by which three chains associate to form triple helical protomers in a chain specific manner. This also plays a major role in the supramolecular assembly of collagen type IV in the extracellular matrix. Tissue specific protomer types of collagen IV exist in mammals, with alpha1.alpha1.alpha2 protomer being ubiquitous and alpha3.alpha4.alpha5 protomer in kidney. The long standing interest of our lab in this project is to determine the crystal structures of NC1 domains in hexameric forms as well as their monomeric components to understand the chain specific assembly type IV collagen. These structures will have strong implications in understanding the role of type IV collagen in tissue development, more specifically, kidney. At least two kidney diseases, namely Alport?s syndrome and Goodpasture?s syndrome, are directly linked to type IV collagen. The molecular structures of [alpha3.alpha4.alpha5]2 and alpha3 monomer are especially crucial in understanding the molecular basis for the pathogenesis of Goodpasture syndrome. This is an autoimmune disease in which the antibodies specifically target alpha3 NC1 domain in the monomer, but not when it is a part of [alpha3.alpha4.alpha5]2 hexamer. We have determined the first crystal structure of NC1 hexamer, [alpha1.alpha1.alpha2]2 using Br-MAD data collected at SSRL. We have also collected data on alpha2 and alpha3 monomer crystals, whose structures could not be solved by molecular replacement method using the monomer models from the hexamer. Next, we would attempt to solve these structures by MAD using Se or Br as anomalous scatterer. We have also obtained weakly diffracting crystals of [alpha3.alpha4.alpha5]2 NC1 hexamer, which could not be characterized using the in-house source.

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。 IV型胶原蛋白是所有后生动物基底膜的主要结构蛋白成分。它包含一个大的C末端球状结构域（NC1结构域），通过该结构域，三个链以特定方式以链的特定方式形成三重螺旋原型。这在细胞外基质中的胶原蛋白IV型超分子组件中也起着重要作用。哺乳动物中存在组织特异性胶原蛋白IV的特定类型类型，α1.Alpha1.Alpha2brotomer无处不在，alpha3.alpha3.alpha4.alpha4 protomer在肾脏中。我们实验室在该项目中的长期兴趣是确定六聚体形式中NC1域的晶体结构以及其单体组件，以了解链式组件类型IV胶原蛋白。这些结构将对理解IV型胶原蛋白在组织发育中的作用（更具体地说是肾脏）具有很大的影响。至少有两种肾脏疾病，即Alport的综合征和Goodpasture综合征，直接与IV型胶原蛋白有关。 [alpha3.alpha4.alpha5] 2和alpha3单体的分子结构对于理解Goodpasture综合征发病的分子基础特别重要。这是一种自身免疫性疾病，其中抗体特异性靶向单体中的alpha3 NC1结构域，而不是当它是[alpha3.alpha4.alpha5] 2的一部分时。我们已经使用SSRL收集的BR-MAD数据确定了NC1六聚体的第一个晶体结构[alpha1.alpha1.alpha2]​​ 2。我们还收集了有关α2和alpha3单体晶体的数据，它们的结构无法通过使用己酰胺的单体模型通过分子替换方法来溶液。接下来，我们将尝试通过使用SE或BR作为异常散射器来解决这些结构。 我们还获得了[alpha3.alpha4.alpha5] 2 NC1六聚体的弱衍射晶体，使用内部源无法表征。