MOLECULAR STRUCTURE, FUNCTION AND PATHOLOGY OF ELASTIN

弹性蛋白的分子结构、功能和病理学

• 批准号: 3340701
• 负责人: DAN W URRY
• 金额: $ 20.8万
• 依托单位: UNIVERSITY OF ALABAMA AT BIRMINGHAM
• 依托单位国家: 美国
• 财政年份: 1982
• 资助国家: 美国
• 起止时间: 1982-05-01 至 1987-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3340701
• 关键词: X ray crystallography; biophysics; chemical structure; chemical synthesis; cholesterol; circular dichroism; computer graphics /printing; conformation; crosslink; elastases; electron microscopy; fibrogenesis; hydroxylation; infrared spectrometry; nuclear magnetic resonance spectroscopy; phase change; radiotracer; relaxation spectrometry; ultraviolet spectrometry; viscosity

The specific aims of this research proposal are: 1) to synthesize component peptides of the elastic fiber, i.e. the repeat peptides, actual sequences containing the repeats, multicomponent models comprised of the repeats, and composities approaching tropoelastin, and 2) to develop three dimensional structures for the peptides for the purpose of understanding the molecular structure, function and pathology of the elastic fiber, utilizing the disciplines of molecular biophysics and pathobiological chemistry. The approach is to study a series of relevant molecular systems of graded, increasing complexity and to do so in each of three states, solution, coacervate and fibrous, which constitute a set of states of graded, increasing complexity. The chemical methods to be used are those of solution and solid phase peptide synthesis. The physical and computational characterizations utilize proton and carbon-13 NMR (including solid state), nuclear Overhauser effects, relaxation studies, CD and UV spectroscopies, and X-ray diffraction in a combined effort to place conformational constrains on the molecular systems and to use conformational energy analyses for further constraints in a stepwise manner to develop descriptions of preferred conformations. These data, at the Beta-spiral and twisted filament level of development, will be combined with ultrastructural characterizations of microscopy (light, scanning and conventional with negative staining) and molecular graphics to obtain a consistent and detailed description of the filamentous structures comprising the fibrous states. The methods utilized in studying the molecular pathology of each of the relevant molecular systems are: calcium-43 NMR relaxation studies to characterize calcium binding, columns made of the fibrous states to study lipid binding, serum dialysate calcification (characterized by SEM and microanalysis), and prolyl hydroxylation as it relates to fiber formation and to degradation by elastase. The elastic fiber is a primary site for the molecular pathologies of calcification and lipid deposition, and is implicated in such disease processes as atherosclerosis, pulmonary emphysema, solar elastosis, and other complications of the connective tissue which are relevant to heart and lung diseases and to aging processes.

本研究的具体目标是：1）综合 弹性纤维的组分肽，即重复肽，实际上 包含重复序列的多组分模型， 重复，和复合物接近原弹性蛋白，和2）开发三个 为了理解肽的三维结构 弹性纤维的分子结构、功能和病理， 利用分子生物物理学和病理生物学的学科， 化学. 方法是研究一系列相关的分子系统 逐渐增加的复杂性，并在三个州中的每一个州这样做， 溶液、凝聚层和纤维层，它们构成一组状态， 分级的，不断增加的复杂性。 要使用的化学方法是 溶液和固相肽合成的最佳方法。 物理和 计算表征利用质子和碳-13 NMR（包括 固态），核Overhauser效应，弛豫研究，CD和UV 光谱学和X射线衍射的结合， 分子系统的构象约束，并使用 构象能分析，以逐步的方式进一步限制 来描述优选的构象。 这些数据，在 β-螺旋丝和加捻丝的发展水平，将结合起来 用显微镜（光，扫描和 常规负染色）和分子图形，以获得 对丝状结构的一致和详细的描述 包括纤维状状态。 研究中使用的方法 每个相关分子系统的分子病理学是： 钙-43 NMR弛豫研究，以表征钙结合，色谱柱 制成的纤维状态，以研究脂质结合，血清透析液 钙化（通过SEM和微量分析表征）和脯氨酰 羟基化，因为它涉及纤维形成和降解， 弹性蛋白酶 弹性纤维是分子的主要场所， 钙化和脂质沉积的病理学，并涉及 这些疾病过程如动脉粥样硬化、肺气肿、日光性 弹性组织变性和其他结缔组织并发症， 与心脏和肺部疾病以及衰老过程有关。