Cattani Julia, Subramaniam Vinod, Drescher Malte
Department of Chemistry and Konstanz Research School Chemical Biology, University of Konstanz, 78457 Konstanz, Germany.
Phys Chem Chem Phys. 2017 Jul 19;19(28):18147-18151. doi: 10.1039/c7cp03432f.
Human alpha-Synuclein (aS), implicated in Parkinson's disease, adopts a rich variety of different conformations depending on the macromolecular context. In order to unravel its pathophysiological role, monitoring its intracellular conformational state and identifying differences for the disease variants is crucial. Here, we present an intracellular spectroscopy approach based on a systematic spin-labeling site-scan in combination with intracellular electron paramagnetic resonance spectroscopy determining conformations on a molecular scale. A quantitative and model-based data analysis revealed that the vast majority of aS, be it wild-type or disease variants A30P or A53T, exists in the monomeric intrinsically disordered form in the cell.
与帕金森病相关的人类α-突触核蛋白（aS），会根据大分子环境呈现出丰富多样的不同构象。为了阐明其病理生理作用，监测其细胞内构象状态并识别疾病变体之间的差异至关重要。在此，我们提出一种基于系统自旋标记位点扫描与细胞内电子顺磁共振光谱相结合的细胞内光谱方法，可在分子尺度上确定构象。基于模型的定量数据分析表明，无论是野生型还是疾病变体A30P或A53T，绝大多数aS在细胞中以单体内在无序形式存在。
