CHEMISTRY/BIOLOGY OF THE GENETICALLY DISTINCT COLLAGENS

这一修订后的竞争性延续角色申请的长期目标 是为了理解生物多样性和存在的功能基础 脊椎动物体内独特的纤维形成胶原系统。这 研究解决了两个普遍的问题；第一，为什么有这么多 形成纤维的胶原蛋白类型；第二，为什么每种胶原蛋白的分布 类型是特定于组织的。本应用重点是III型胶原蛋白AS 一个模型系统，以确定 这种胶原蛋白在人体不同组织中的独特分布。这个 首席调查员最近描述了一种快速而敏感的 高效液相色谱分离技术的研究进展 拆分和定量每种胶原蛋白的一个小标记多肽。 具体的目标是：第一，检验比例的假设 在可膨胀的组织中，III型胶原的含量增加。最近的 所描述的高效液相技术将用于确定类型的比例 人胸主动脉内特定区域的I型和III型胶原(高度 可扩张的)和腹主动脉(不太顺应性)来自年轻男子 还有女人。该方法依赖于字母(I)-CB2的检测， Alphal(III)-CB2和Alphal(V)-CBI氰基溴化肽。类型 高度扩张的人胸主动脉和胸主动脉的III型胶原含量 不符合条件的大动脉样本将被确定。这项研究应该测试 关于III型胶原在膨胀性疾病中更为普遍的假说 纸巾。第二个具体目标是确定类型III的作用 胶原蛋白作为主要贡献者，与II型胶原蛋白一起， 不可膨胀组织的生物学功能。高效液相色谱仪将用于 开发一种可比较的分析系统来确定 透明软骨标本中形成纤维的胶原蛋白，即II，III， IX和XI胶原蛋白。第三个具体目标是检验这一假设 III型胶原的主要替代作用是促进 一种名为短肽的新型丝氨酸蛋白酶抑制剂的结合 α-L-抗胰蛋白酶对细胞外基质的作用。此装订 至少使III型胶原，可能还有其他大分子，能够 抵抗丝氨酸蛋白酶的不适当攻击。的职能作用 SPAAT与III型胶原的优先结合将是 通过评估SPAAT活性片段的能力来确定 抑制III型胶原降解的纤维形成胶原 纤维在体外形成。第四个具体目标是测试 假设SPAAT将主要定位于类型丰富的组织 III型胶原。SPAAT的组织分布将通过以下方式评估 使用特定抗体的免疫组织化学技术。

The long-term goal of this revised competing continuation ROl application is to understand the functional basis for the diversity and presence of the unique fiber-forming collagen systems in vertebrate organisms. This research addresses two general questions; first, why there are so many fiber-forming collagen types; and second, why the distribution of each type is tissue-specific. This application focuses on type III collagen as a model system to determine the structural and functional basis for the unique distribution of this collagen in different human tissues. The Principal Investigator has recently described a rapid and sensitive technique relying on high pressure liquid chromatography (HPLC) for the resolution and quantitation of a small marker peptide for each collagen. The Specific Aims are: first, to test the hypothesis that the proportions of type III collagen are elevated in distensible tissues. The recently described HPLC technique will be used to determine the proportions of type I and III collagens from defined regions in human thoracic aorta (highly distensible) and abdominal aorta (less compliant) derived from young men and women. The method depends on the detection of the alphal(I)-CB2, alphal(III)- CB2, and alphal(V)-CBI cyanogen bromide peptides. The type III collagen content of highly distensible human thoracic aorta and noncompliant aorta samples will be determined. This study should test the hypothesis that type III collagen is more prevalent in distensible tissues. The second specific aim is to establish the role of type III collagen as a major contributor, along with type II collagen, to the biological function of a nondistensible tissue. HPLC will be used to develop a comparable analytical system for determining the proportions of fiber-forming collagens in hyaline cartilage samples, i.e., type II, III, IX and XI collagens. The third specific aim is to test the hypothesis that a major alternative role of type III collagen is to facilitate binding of a novel inhibitor of serine proteases termed short peptide from alpha l-antitrypsin (SPAAT) to extracellular matrices. This binding enables at least type III collagen, and possibly other macromolecules, to resist inappropriate attack by serine proteases. The functional role of the preferential binding of a SPAAT to type III collagen will be determined by evaluating the ability of the active fragment of SPAAT to the fiber-forming collagens to inhibit degradation of type III collagen fibrils formed in vitro. The fourth specific aim is to test the hypothesis that SPAAT will be localized largely in tissues rich in type III collagen.Tissue distribution of SPAAT will be evaluated by immunohistochemical techniques using specific antibodies.