来自与疾病相关的氨酸扩张域的纤维的结构和动态表征,使用蛋白质分解和固态NMR光谱学
Mirko Sackewitz1, Holger A Scheidt, Grit Lodderstedt
1Institute of Biochemistry and Biotechnology, Martin Luther University Halle-Wittenberg, D-06120 Halle, Germany.
Journal of the American Chemical Society
|May 17, 2008
概括
核蛋白 PABPN1 的核蛋白.
科学领域:
- 生物化学 生物化学
- 分子生物学分子生物学
- 生物物理学的生物物理.
背景情况:
- 眼肌肉发育不良 (OPMD) 与核聚A结合蛋白1 (PABPN1) 中扩展的氨酸重复有关.
- 这些扩张导致核内纤维状沉积物,这是OPMD的一个关键本病学特征.
研究的目的:
- 为了研究PABPN1的纤维状N端片段的结构和动态特性,具有扩展的氨酸延伸 (N-(+7) Ala).
- 了解聚氨酸延伸在纤维细胞形成和蛋白质动态中的作用.
主要方法:
- 固态NMR光谱 (13C MAS NMR) 用于分析蛋白质结构和动态.
- 蛋白质分解裂变和MALDI-TOF质谱学确定了蛋白质酶耐药的区域.
- 对1H-13Calpha二极合物的特定场所测量确定了顺序参数.
主要成果:
- 交叉极化13C MAS NMR显示了氨酸残留的特征性β-sheet信号.
- 直接偏光的光谱揭示了蛋白质中高度移动的区域.
- 确定了一种含有聚氨酸延伸的蛋白酶耐药核心.
- 聚氨酸延伸显示了高阶参数 (0.77),表明了刚性纤维结构.
- 其他N-(+7) Ala的区域显示低级参数 (0.06-0.15),表明高流动性,除了侧面的Gly残留物 (0.47).
结论:
- 在PABPN1中的聚氨酸延伸形成了一个高度有序的纤维状结构.
- 使用NMR的特定站点动态映射对于表征纤维状蛋白的拓学是有效的.
- 这些发现提供了对OPMD病原体背后的分子机制的见解.
更多相关视频
12:58Characterizing Individual Protein Aggregates by Infrared Nanospectroscopy and Atomic Force Microscopy
Published on: September 12, 2019
07:56Utilizing Time-Resolved Protein-Induced Fluorescence Enhancement to Identify Stable Local Conformations One α-Synuclein Monomer at a Time
Published on: May 30, 2021
相关概念视频
03:03Amyloid Fibrils
Amyloid fibrils are aggregates of misfolded proteins. Under most circumstances, misfolded proteins are either refolded by chaperone proteins or degraded by the proteasome. However, in the case of a mutation or a disease, these proteins can accumulate to form large clusters and often further assemble to form elongated fibers, called fibrils.
Amyloid deposits were observed as early as 1639 in the liver and the spleen. In 1854, Rudolph Virchow performed iodine staining, normally used to...
Amyloid deposits were observed as early as 1639 in the liver and the spleen. In 1854, Rudolph Virchow performed iodine staining, normally used to...
03:03Amyloid Fibrils
Amyloid fibrils are aggregates of misfolded proteins. Under most circumstances, misfolded proteins are either refolded by chaperone proteins or degraded by the proteasome. However, in the case of a mutation or a disease, these proteins can accumulate to form large clusters and often further assemble to form elongated fibers, called fibrils.
Amyloid deposits were observed as early as 1639 in the liver and the spleen. In 1854, Rudolph Virchow performed iodine staining, normally used to...
Amyloid deposits were observed as early as 1639 in the liver and the spleen. In 1854, Rudolph Virchow performed iodine staining, normally used to...
01:22Protein Folding
Overview