CHEMISTRY/BIOLOGY OF THE GENETICALLY DISTINCT COLLAGENS

这一修订后的竞争性继续ROI申请的长期目标是 是了解的多样性和存在的功能基础， 脊椎动物中独特的纤维形成胶原系统。 这 研究解决了两个一般性问题;第一，为什么有这么多 纤维形成胶原蛋白类型;第二，为什么每种类型的分布 类型是组织特异性的。 本申请集中于III型胶原蛋白， 一个模型系统，以确定结构和功能的基础， 这种胶原蛋白在不同人体组织中的独特分布。 的 首席研究员最近描述了一种快速而敏感的 技术依赖于高压液相色谱法（HPLC）， 每种胶原蛋白的小标记肽的解析和定量。 具体目的是：首先，检验比例 III型胶原蛋白在可扩张组织中升高。 最近 将使用所描述的HPLC技术来确定类型 I和III胶原蛋白从人胸主动脉的定义区域（高度 可扩张）和腹主动脉（顺应性较差）来自年轻男性 和妇女 该方法依赖于对双醛（I）-CB 2的检测， β-半乳糖醛（III）-CB 2和β-半乳糖醛（V）-CBI溴化氰肽。 的类型 高度扩张的人胸主动脉的III型胶原含量和 将确定不顺应性主动脉样本。 这项研究将测试 假设III型胶原蛋白在可扩张性组织中更普遍， 组织中 第二个具体目标是确立第三类的作用 胶原作为主要贡献者，沿着II型胶原， 不可扩张组织的生物学功能。 HPLC将用于 开发一个可比较的分析系统，用于确定 透明软骨样品中的纤维形成胶原，即，II型，III型， IX和Xi胶原蛋白。 第三个具体目标是检验假设 III型胶原蛋白的主要替代作用是促进 一种称为短肽的丝氨酸蛋白酶的新型抑制剂与来自 α l-抗胰蛋白酶（SPAAT）与细胞外基质的结合。 此绑定 使至少III型胶原蛋白和可能的其它大分子， 抵抗丝氨酸蛋白酶的不适当攻击。 的功能作用 SPAAT与III型胶原的优先结合将是 通过评估SPAAT的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， SPAAT的组织分布将通过 免疫组织化学技术使用特异性抗体。

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.