Fiber diffraction of synthetic α-synuclein filaments shows amyloid-like cross-β conformation
Fiber diffraction of synthetic α-synuclein filaments shows amyloid-like cross-β conformation
复制标题
DOI：
10.1073/pnas.97.9.4897
复制
发表时间：
2000-04-25
影响因子：
11.1
通讯作者：
Crowther, RA
中科院分区：
文献类型：
--
作者：
Serpell, LC;Berriman, J;Crowther, RA
由α-突触核蛋白组成的丝状包涵体构成了帕金森病、路易体痴呆和多系统萎缩的神经病理学特征。罕见的帕金森病家族病例与 α-突触核蛋白中的 A53T 和 A30P 突变有关。我们在此报告重组α-突触核蛋白的组装特性和二级结构特征。羧基末端截短的人 α-突触核蛋白 (1-87) 和 (1-120) 显示出最快的组装速度，其次是人 A53T α-突触核蛋白，以及大鼠和斑胸草雀 α-突触核蛋白。野生型人类 α-突触核蛋白和 A30P 突变体表现出较慢的组装速度。摇动后，在 37 摄氏度下 48 小时内形成细丝。相关蛋白 β- 和 γ-突触核蛋白仅在孵育几周后才组装。合成的人 α-突触核蛋白丝由针对 α-突触核蛋白羧基端 10 个氨基酸的抗体修饰，从路易体痴呆症和多系统萎缩脑中提取的丝也是如此。圆二色光谱表明，α-突触核蛋白在组装过程中经历了从无规卷曲到β-折叠结构的构象变化。 α-突触核蛋白组装体的 X 射线衍射和电子衍射显示淀粉样蛋白的交叉 β 构象特征。
Filamentous inclusions made of alpha-synuclein constitute the defining neuropathological characteristic of Parkinson's disease, dementia with Lewy bodies, and multiple system atrophy. Rare familial cases of Parkinson's disease are associated with mutations A53T and A30P in alpha-synuclein. We report here the assembly properties and secondary structure characteristics of recombinant alpha-synuclein. Carboxy-terminally truncated human alpha-synuclein (1-87) and (1-120) showed the fastest rates of assembly, followed by human A53T alpha-synuclein, and rat and zebra finch alpha-synuclein. Wild-type human alpha-synuclein and the A30P mutant showed slower rates of assembly. Upon shaking, filaments formed within 48 h at 37 degrees C. The related proteins beta- and gamma-synuclein only assembled after several weeks of incubation. Synthetic human alpha-synuclein filaments were decorated by an antibody directed against the carboxy-terminal 10 amino acids of alpha-synuclein, as were filaments extracted from dementia with Lewy bodies and multiple system atrophy brains. Circular dichroism spectroscopy indicated that alpha-synuclein undergoes a conformational change from random coil to beta-sheet structure during assembly. X-ray diffraction and electron diffraction of the alpha-synuclein assemblies showed a cross-beta conformation characteristic of amyloid.