PROBING THE BIOCHEMICAL MECHANISM OF AMYLOID DISEASES

探索淀粉样蛋白疾病的生化机制

• 批准号: 2905537
• 负责人: JEFFERY W KELLY
• 金额: $ 26.47万
• 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/2905537
• 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融资后的第一次竞争性续约 探讨淀粉样变性疾病的发病机制。这样做的长期目标是 研究计划是为了了解人类的生化机制 淀粉样变性和开发新的治疗策略来验证假说 关于这些疾病的病因学。 人类淀粉样变性疾病的共同特征涉及细胞外 蛋白质沉积在纤维形态中。重大间接证据 有证据表明，纤维直接导致了 大脑相关疾病，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人员将他的建议集中在两个淀粉样蛋白上 不直接累及大脑的疾病。这些是家族性淀粉样蛋白 多发性神经病和老年性系统性淀粉样变性。在家族性疾病中， 突变形式的反式甲状腺激素(前白蛋白)(TTR)在 周围神经或特定器官，而在衰老的形式 疾病正常的TTR主要形成于心脏组织。这些疾病是 选择用来检验普通淀粉样蛋白假说的小分子 可抑制TTR淀粉样原纤维形成的分子直接评估 抑制转甲状腺素原纤维的形成是否足以 预防淀粉样变性疾病的发病。 这项建议的第一个具体目标是围绕进一步的测试 构象变化假说，它表明第三纪 结构变化涉及到使一种特定蛋白质产生淀粉样蛋白。从… 先前的工作表明，TTR淀粉样原纤维的形成是由 蛋白质的另一种三级结构的自组装。这 结果将在继续资助的情况下得到扩展，以应用大量的生物物理 包括质谱学和核磁共振法在内的精确测定方法 野生型淀粉样变性中间体的结构，并比较这一点 家族性淀粉样多发性神经病相关变异体的结构 转甲状腺素(TtR)。过去资助期内的先前工作表明 这些Ttr变异体的稳定性要差得多，变性程度要高出一千倍 比野生型Ttr更快。在持续的支持下，凯利博士将 研究由野生型和家族性混合变异组成的四聚体 评估它们的稳定性、pH介导的变性速率和 淀粉样变性。 第二个目标涉及基于结构的药物设计策略，该策略利用 X-射线结晶学、有机合成和有机合成的协同应用 用于鉴定高亲和力结合淀粉样蛋白的平行筛选 稳定Ttr天然状态并防止pH下降的抑制剂 介导的导致纤维形成的构象变化。