CRYSTAL STRUCTURE DETERMINATION OF COLLAGEN-RELATED STRUCTURES

胶原相关结构的晶体结构测定

• 批准号: 7370482
• 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: 2006
• 资助国家: 美国
• 起止时间: 2006-03-01 至 2007-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7370482
• 关键词: CRYSTAL; STRUCTURE; DETERMINATION; COLLAGEN; RELATED

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 ?1.?1.?2 protomer being ubiquitous and ?3.?4.?5 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 [?3.?4.?5]2 and ?3 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 ?3 NC1 domain in the monomer, but not when it is a part of [?3.?4.?5]2 hexamer. We have determined the first crystal structure of NC1 hexamer, [?1.?1.?2]2 using Br-MAD data collected at SSRL. We have also collected data on ?2 and ?3 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 [?3.?4.?5]2 NC1 hexamer, which could not be characterized using the in-house source.

这个子项目是利用由NIH/NCRR资助的中心拨款提供的资源的许多研究子项目之一。子项目和调查员(PI)可能从另一个NIH来源获得了主要资金，因此可能会出现在其他CRISE条目中。列出的机构是针对中心的，而不一定是针对调查员的机构。IV型胶原是所有后生动物基底膜的主要结构蛋白成分。它含有一个大的C-末端球状结构域(NC1域)，通过它，三条链以链特异的方式结合形成三个螺旋原基。这也在细胞外基质中IV型胶原的超分子组装中发挥着重要作用。在哺乳动物中存在组织特异性的IV型胶原原基类型，其中？1.？1.？2原基在肾脏中普遍存在，？3.4.？5原基存在于肾脏中。我们实验室长期以来对这个项目的兴趣是确定六聚体形式的NC1结构域的晶体结构以及它们的单体组成，以了解链特异性组装的IV型胶原。这些结构将对理解IV型胶原在组织发育中的作用，更具体地说，在肾脏中具有重要意义。至少有两种肾脏疾病，即阿尔波特S综合征和好牧场S综合征，与IV型胶原直接相关。[？3.？4.？5]2和？3单体的分子结构对于了解古巴德综合征发病的分子基础尤为重要。这是一种自身免疫性疾病，抗体特异性地针对单体中的？3NC1域，但当它是[？3.？4.？5]2六聚体的一部分时不是。我们用SSRL收集的BR-MAD数据确定了NCl六聚体的第一个晶体结构[？1.？1.？2]2。我们还收集了？2和？3单体晶体的数据，它们的结构不能用分子置换方法从六聚体的单体模型中计算出来。下一步，我们将尝试用Se或Br作为反常散射体来解决这些结构。我们还获得了[？3.4.？5]2NCl六角体的弱衍射晶体，这是用内部光源无法表征的。