Laboratory of Molecular Biophysics, National Heart, Lung, and Blood Institute, National Institutes of Health, Bethesda, Maryland 20892, United States.
Biochemistry. 2011 Mar 29;50(12):1963-5. doi: 10.1021/bi2000824. Epub 2011 Feb 21.
In the Parkinson's disease-associated state, α-synuclein undergoes large conformational changes, forming ordered, β-sheet-containing fibrils. To unravel the role of specific residues during the fibril assembly process, we prepared single-Cys mutants in the disordered (G7C and Y136C) and proximal (V26C and L100C) fibril core sites and derivatized them with environmentally sensitive dansyl (Dns) fluorophores. Dns fluorescence exhibits residue specificity in spectroscopic properties as well as kinetic behavior; early kinetic events were revealed by probes located at positions 7 and 136 compared to those at positions 26 and 100.
在帕金森病相关状态下，α-突触核蛋白发生大的构象变化，形成有序的、含β-折叠的纤维。为了揭示特定残基在纤维组装过程中的作用，我们制备了无规卷曲（G7C 和 Y136C）和近纤维核心（V26C 和 L100C）位点的单-Cys 突变体，并将其衍生化，用环境敏感的丹磺酰（Dns）荧光团进行标记。Dns 荧光在光谱特性和动力学行为上都表现出残基特异性；与位于位置 26 和 100 的探针相比，位于位置 7 和 136 的探针揭示了早期动力学事件。
