Development of a high-pressure treatment for amyloidosis

淀粉样变性高压治疗方法的开发

• 批准号: 13558082
• 负责人: TACHIBANA Hideki
• 金额: $ 7.62万
• 依托单位: Kobe University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (B)
• 财政年份: 2001
• 资助国家: 日本
• 起止时间: 2001 至 2003
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-13558082/
• 关键词: lysozyme; amyloid; high-pressure NMR; prion; metastable intermediate; 国際情報交換; 米国; アミロイド病; βラクトグロブリン; lysozyme; amyloid; high-pressure NMR; prion; metastable intermediate; 国際情報交換; 米国; アミロイド病; βラクトグロブリン

1. A disulfide-deficient variant of hen lysozyme self-associates into a 17-S assemblage that contains intermolecular beta-structure. Upon prolonged incubation over weeks to months, amyloid-like fibrils are spontaneously formed. No specific, partially unfolded conformer is involved, and fibrils are formed starting from an intrinsically unfolded monomeric state and passing through a precursor with a spontaneous build-up of intermolecular beta-structure.2. The 17-S precursor reversibly dissociates under hydrostatic pressure of several hundred to two thousand bars. It is characterized with a Gibbs energy for association of -23.3 kJ and a volume increase of 52.7 ml per mol of monomer unit, and involves interaction of hydrophobic residues in the initial association step.3. By carrying out two-dimensional NMR measurements under variable pressure, a metastable conformer of the normal cellular isoform of prion protein (PrP^c) has been found to occur at a population of 1 %, in which helices B an … More d C are disordered. This metastable conformer is most logically PrP^* that is implicated in the step for transformation of PrP^c to the infectious prion protein (PrP^<Sc>) via interaction with a template PrP^<Sc>. On the other hand, the conformational characteristics of a fibril-forming fragment of the mouse prion protein have been investigated by using hydrogen-deuterium exchange measurements and molecular dynamics simulations. Fibril formation was shown to involve polyalanine stacking.4. There exists a correlation for prion protein between the residue-wise stability determined by high-pressure NMR spectroscopy and slow fluctuations determined by the CPMG method at ambient pressure. This indicates that the metastable conformer revealed by high-pressure NMR can be acquired through slow conformational fluctuations under physiological conditions. Cavities exist at the site of large fluctuation, which strongly suggests that the screening for the molecules that bind to the cavity leads to the development of drugs for the treatment of prion disease or amyloidosis. Less

1. HEN溶菌酶自我缔合的二硫键缺陷变体成17-S组合，其中包含分子间β结构。在长时间孵育数周到几个月的时间内，赞助了淀粉样蛋白样的原纤维。不涉及特定的，部分展开的构象体，并从本质上展开的单体状态开始并通过具有共分子间β结构的堆积的前体形成原纤维。2。 17-S前体在静水压力下可逆分解几百至两千个棒。它的特征是吉布斯的能量，用于-23.3 kJ的缔合和每摩尔单体单位的体积增加52.7 mL，并且涉及在初始关联步骤中疏水保留的相互作用。3。通过在可变压力下进行二维NMR测量，发现正常细胞同工型（PRP^C）的亚稳态构象异构体发生在1％的人群中，其中螺旋螺旋b和更多的D C是无序的。这种亚稳态的构象异构体是最逻辑上的prp^*，它是通过与模板prp^<sc>的相互作用的相互作用在PRP^c转换为感染性prion蛋白（PRP^<sc <sc）的步骤中实现的。另一方面，已经通过使用氢 - 脱氧群交换测量和分子动力学模拟研究了小鼠prion蛋白纤维形成片段的构象特性。纤维形成显示涉及多酰胺堆积4。通过高压NMR光谱法确定的居住稳定性和在环境压力下通过CPMG方法确定的缓慢波动之间存在的主要蛋白质相关。这表明可以通过生理条件下缓慢的构象波动来获取高压NMR揭示的亚稳态构象异构体。空腔存在于大波动的部位，这强烈表明筛选与腔内结合的分子的筛查会导致药物的开发，以治疗治疗prion病或淀粉样变性。较少的