STRUCTURE AND ASSEMBLY OF COLLAGEN MOLECULES AND FIBRILS

胶原蛋白分子和原纤维的结构和组装

• 批准号: 3154794
• 负责人: ARTHUR VEIS
• 金额: $ 18.96万
• 依托单位: NORTHWESTERN UNIVERSITY AT CHICAGO
• 依托单位国家: 美国
• 财政年份: 1979
• 资助国家: 美国
• 起止时间: 1979-03-01 至 1989-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3154794
• 关键词: acidity /alkalinity; basement membrane; chemical aggregate; conformation; connective tissue metabolism; endoplasmic reticulum; exopeptidase; fibroblasts; high performance liquid chromatography; immunochemistry; lens; polysomes; proteoglycan; tissue /cell culture

Collagen molecules are the principal structural elements of all connective tissues. Collagen fiber organization and ultrastructure varies from one tissue to the next. The objective of this study is to delineate the basic molecular events which are involved in the molecular fibril assembly processes and to determine the factors which regulate these processes. Four specific aspects of assembly are to be examined in detail. 1) Nascent pro Alpha chains register in the appropriate stoichiometry and initiate triple helix formation as the chains near release from the endoplasmic reticulum membrane. Membrane mediated interactions may guide this phase of assembly. A combination of cell-free translation, polysome isolation and completion, and membrane insertion studies will be used to examine the chain assembly mechanism. 2) Interstitial procollagens must be processed at both COOH - and NH2-propeptides before stable fibrils form. The NH2-propeptidase activity is well understood whereas the COOH- propeptidase has not been isolated and the site of its action not determined. Evidence is accumulating that the COOH-propeptide cleavage might be tied in with late stages of intracellular secretory processing and regulation of intracellular degradation. The processing localization of this event, and its enzymology are to be examined in detail. 3) Fibril formation is an event dominated by telopeptide-helix receptor region interactions. The explicit chemistry of this interaction, and the properties of these molecular domains are to be examined by interaction studies between isolated peptides, by physical studies (CD, FT-IR), and by interaction and conformational predictive studies. 4) Type IV basement membrane collagens form filamentous networks rather than fibrils, from molecules which are minimally processed and retain extensive non-triplet sequence chain segments. Anterior lens-capsule Type IV collagen can be obtained with minimal degradation. The self-assembly process will be examined, by thermal self-assembly studies and various electron microscopic techniques including SLS formation and rotary shadowing. From all these studies we should develop better insights into the synthesis and construction of the collagenous framework of the connective tissues. Our studies of the mechanism of molecular assembly and tissue ordering have the ultimate objective of providing insight into the control of the connective tissue disorders and their treatment.

胶原蛋白分子是所有结缔组织的主要结构元素 纸巾。胶原纤维组织和超微结构千差万别 纸巾传给下一个。这项研究的目的是勾勒出基本的 参与分子原纤维组装的分子事件 并确定规范这些过程的因素。 将详细研究组装的四个具体方面。1)萌芽状态 Pro Alpha链在适当的化学计量比中注册并启动 当链接近从内质中释放时，形成三螺旋结构 网状膜。膜介导的相互作用可能指导这一阶段的 集合。无细胞翻译、多聚体分离和 完成，并将使用膜插入研究来检查 链条装配机构。2)必须处理间质前胶原 在稳定的纤维形成之前，在COOH-和NH2-前肽。这个 NH2-前肽酶活性是众所周知的，而COOH-前肽酶 没有被孤立，其行动地点也没有确定。证据 COOH-前肽的裂解可能与 细胞内分泌过程的晚期及其调控 细胞内降解。此事件的处理本地化，以及 将对其进行详细的酶学检查。3)原纤层是一种 端肽-螺旋受体区域相互作用主导的事件。这个 这种相互作用的显性化学，以及它们的性质 分子结构域将通过相互作用研究来检验 通过物理研究(CD，FT-IR)和通过相互作用和 构象预测研究。4)IV型基底膜胶原 形成丝状网络而不是纤维，分子是 最小限度地加工并保留广泛的非三联序列链 细分市场。晶状体前囊膜IV型胶原蛋白的制备 最小降级。将通过以下方式检查自组装过程 热自组装研究和各种电子显微镜技术 包括SLS编队和旋转阴影。从所有这些研究来看，我们 应该更好地洞察 结缔组织的胶原性骨架。我们对人类社会的研究 分子组装和组织有序的机制具有终极意义 目的为结缔组织的控制提供洞察力 疾病及其治疗。