Kinetic and thermodynamic analysis of the molecular interaction on amyloid fibril formation

淀粉样蛋白是蛋白质的不溶性纤维状聚集体的细胞外存款，这些蛋白质在其天然构象下通常是可溶的，导致组织结构和功能的损害。我们已经建立了几种体外淀粉样纤维形成系统，包括阿尔茨海默病的β-淀粉样蛋白（Aβ）和透析相关淀粉样变性的β2-微球蛋白（β2-m）淀粉样蛋白（Aβ 2 M）。利用这些系统，发现了各种生物分子或化合物对淀粉样纤维的形成和解离的重要作用。为了通过体外实验系统或数值模型模拟体内淀粉样纤维的形成，我们选择了纤维形成或解离的几个阶段，并通过生物物理、动力学和热力学分析阐明了它们的机制.为了了解Aβ 2-M淀粉样变性的分子发病机制，需要了解通过影响β2-m和淀粉样纤维的构象和稳定性来诱导淀粉样纤维形成的生物分子。 关于我们 ntified.我们发现，阴离子两亲性化合物，包括非酯化脂肪酸（NEFAs）和一些溶血磷脂诱导的原纤维延伸在中性pH值。此外，血液透析患者的溶血磷脂的血浆浓度显着高于健康受试者。这些结果表明溶血磷脂和NEFA在Aβ 2 M淀粉样变性的发展中可能起作用。采用分光光度法、荧光分光光度法和表面等离子体共振技术，研究了5种黄酮类化合物在体外对β-淀粉样纤维（fAβ）的抗淀粉样变性作用。结果表明，黄酮类化合物，尤其是杨梅素，在体外通过优先可逆地结合fAβ的淀粉样纤维结构而不是Aβ单体发挥抗淀粉样蛋白的作用.我们开发了一种高通量筛选系统，用于使用硫磺素T荧光光谱法进行Aβ 2 M和Aβ淀粉样纤维的成核。利用该系统，研究了各种生物分子包括蛋白多糖、糖胺多糖的作用。本研究在阐明Aβ 2 M和Aβ淀粉样纤维形成机制方面取得了显著进展。少

Amyloid is extracellular deposit of insoluble fibrillar aggregate of proteins those are normally soluble in their native conformation, resulting in the impairment of tissue structure and function. We have established several amyloid fibril formation systems in vitro, including β-amyloid of Alzheimer's disease (Aβ) and β2-microglobulin (β2-m) amyloid (Aβ2M) of dialysis-related amyloidosis. Using these systems, important roles of various biological molecules or compounds on the formation and dissociation of amyloid fibrils were discovered. To model the amyloid fibril formation in vivo by in vitro experimental system or numerical model, we selected several phases of fibril formation or dissociation, and clarified their mechanism by biophysical, kinetic and thermodynamic analyses.1. To understand the molecular pathogenesis of Aβ2M amyloidosis, the biological molecules that induce amyloid fibril formation by affecting the conformation and stability of β2-m and amyloid fibrils need to be ide … More ntified. We showed that anionic amphipathic compounds including non-esterified fatty acids (NEFAs) and some lysophospholipids induce the fibril extension at a neutral pH. Furthermore, hemodialysis patients had significantly higher plasma concentrations of lysophospholipids than healthy subjects. These results suggest possible role of lysophospholipids and NEFAs in the development of Aβ2M amyloidosis.2. Using spectrophotometry, spectrofluorometry and surface plasmon resonance, we investigated the anti-amyloidogenic effects of five flavonoids on β-amyloid fibril (fAβ) in vitro. The results suggest that flavonoids, especially myricetin exert an anti-amyloidogenic effect in vitro by preferentially and reversibly binding to the amyloid fibril structure of fAβ, rather than to Aβ monomers.3. We developed a high-throughput screening system for the nucleation of Aβ2M and Aβ amyloid fibril using thioflavin T fluorescence spectroscopy. Using this system, effect of various biological molecules including proteoglycans, glycosaminoglycans, were investigated. As the conclusion of this study, the remarkable advance in the clarification of the mechanism of Aβ2M and Aβ amyloid fibril formation were obtained. Less