PROBING THE BIOCHEMICAL MECHANISM OF AMYLOID DISEASES

探索淀粉样蛋白疾病的生化机制

• 批准号: 6380781
• 负责人: JEFFERY W KELLY
• 金额: $ 27.83万
• 依托单位: SCRIPPS RESEARCH INSTITUTE, THE
• 依托单位国家: 美国
• 财政年份: 1993
• 资助国家: 美国
• 起止时间: 1993-05-01 至 2002-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6380781
• 关键词: aging; amyloid proteins; amyloidosis; animal genetic material tag; gene mutation; intermolecular interaction; laboratory mouse; molecular assembly /self assembly; neuritic plaques; nuclear magnetic resonance spectroscopy; nucleic acid sequence; pathologic process; protein biosynthesis; retinoid binding proteins; thyroid hormone binding protein; tissue /cell culture

DESCRIPTION: This is the first competitive renewal following R29 funding to investigate the mechanisms of amyloid disease. The long term goals of this research program are to understand the biochemical mechanism of human amyloid disease and develop new therapeutic strategies to test hypotheses about the etiology of these diseases. The common features of human amyloid disease involve the extracellular deposition of proteins in a fiber morphology. Significant circumstantial evidence suggests that fibrils directly cause the neuropathology of the brain associated diseases and this concept has been generalized as the "amyloid hypothesis". This outstanding young investigator focuses his proposal on two amyloid diseases that do not directly involve the brain. These are familial amyloid polyneuropathy and senile systemic amyloidosis. In the familial disease, a mutated form transthyretin (prealbumin) (TTR) is deposited as fibrils in peripheral nerves or in specific organs while in the senile form of the disease normal TTR forms primarily in cardiac tissue. These diseases were chosen in order to test the general amyloid hypothesis by utilizing small molecules that can inhibit TTR amyloid fibril formation to directly assess whether inhibition of transthyretin fibril formation is sufficient to prevent the onset of amyloid disease. The first specific aim of this proposal is centered around further testing the conformational change hypothesis, which suggests that tertiary structural changes are involved to make a given protein amyloidogenic. From prior work, it was shown that TTR amyloid fibril formation results from the self-assembly of an alternative tertiary structure of the protein. This result will be extended with continued funding to apply numerous biophysical methods including mass spectrometry and NMR to precisely determine the structure of the wild type amyloidogenic intermediate, and to compare this structure to the familial amyloid polyneuropathy associated variants of transthyretin (TTR). Previous work during the past funding period has shown that these TTR variants are much less stable and denature a thousand fold faster than wild-type TTR. Under continued support, Dr. Kelly will investigate tetramers composed of mixed wild-type and familial variants to assess their stability, rates of pH mediated denaturation and amyloidogenicity. The second aim involves a structure-based drug design strategy utilizing the synergistic application of x-ray crystallography, organic synthesis and parallel screening in order to identify high affinity binding amyloid inhibitors that stabilize the native state of TTR and prevent the pH mediated conformational changes that result in fibril formation.

描述：这是继R29资助之后的第一次竞争性续约， 研究淀粉样疾病的机制。 这一长期目标 研究计划是了解人类的生化机制， 淀粉样蛋白疾病，并开发新的治疗策略来测试假设 关于这些疾病的病因。 人类淀粉样蛋白疾病的共同特征涉及细胞外 蛋白质以纤维形态沉积。 重要的间接证据 有证据表明，原纤维直接导致神经病理学的 脑相关疾病，这一概念已被概括为 淀粉样蛋白假说 这位杰出的年轻研究者将他的建议集中在两种淀粉样蛋白上 不直接涉及大脑的疾病。 这些是家族性淀粉样蛋白 多发性神经病和老年系统性淀粉样变性。 在家族性疾病中， 突变形式的甲状腺素运载蛋白（前白蛋白）（TTR）作为原纤维沉积在 周围神经或特定器官，而在老年形式的 疾病正常TTR主要在心脏组织中形成。 这些疾病是 选择它是为了通过利用小的 可以抑制TTR淀粉样纤维形成以直接评估的分子 甲状腺素运载蛋白原纤维形成的抑制是否足以 预防淀粉样疾病的发生。 该提案的第一个具体目标是围绕进一步测试 构象变化假说，这表明， 结构变化涉及使给定的蛋白质淀粉样变。 从 先前的工作表明，TTR淀粉样纤维的形成是由 蛋白质的替代三级结构的自组装。 这 结果将继续扩大资金，以应用许多生物物理 包括质谱和NMR在内的方法来精确地确定 野生型淀粉样蛋白生成中间体的结构，并将其 家族性淀粉样多发性神经病相关变异体的结构 甲状腺素运载蛋白（TTR）。 在过去的资助期间， 这些TTR变异体的稳定性要差得多， 比野生型TTR更快。 在持续的支持下，凯利博士将 研究由混合的野生型和家族性变体组成的四聚体， 评估其稳定性、pH值介导的变性率， 淀粉样变性 第二个目标涉及基于结构的药物设计策略， X射线晶体学、有机合成和 平行筛选以鉴定高亲和力结合淀粉样蛋白 稳定TTR的天然状态并防止pH 介导的构象变化导致原纤维形成。