Fluorescence lifetime dynamics of enhanced green fluorescent protein in protein aggregates with expanded polyglutamine
Fluorescence lifetime dynamics of enhanced green fluorescent protein in protein aggregates with expanded polyglutamine
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DOI：
10.1117/1.3290821
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发表时间：
2010-01-01
影响因子：
3.5
通讯作者：
Cheng, Tzu-Hao
中科院分区：
文献类型：
--
作者：
Ghukasyan, Vladimir;Hsu, Chih-Chun;Cheng, Tzu-Hao
蛋白质聚集是许多神经退行性疾病最终导致神经元死亡的特征步骤之一，需要对聚集进行深入研究以开发有效的治疗方法。我们应用荧光寿命成像来表征增强型绿色荧光蛋白（eGFP）与聚q扩展的聚谷氨酰胺拉伸融合后的荧光动力学。在超过39个残基的polyQ扩展处，它具有形成淀粉样蛋白样原纤维和聚集体的固有倾向，并且是亨廷顿氏病的原因。实验结果表明，eGFP与97Q蛋白融合表达会导致eGFP荧光寿命减少约300 ps。这种现象在hsp104缺陷细胞中不会出现，因为hsp104缺陷细胞阻止了polyQ的聚集。我们证明了观察到的寿命减少与聚集本身有关，并讨论了折射率和同质fret在这些动力学中的可能作用。(C) 2010光学仪器工程师学会。(DOI: 10.1117/1.3290821)
Protein aggregation is one of the characteristic steps in a number of neurodegenerative diseases eventually leading to neuronal death and thorough study of aggregation is required for the development of effective therapy. We apply fluorescence lifetime imaging for the characterization of the fluorescence dynamics of the enhanced green fluorescent protein (eGFP) in fusion with the polyQ-expanded polyglutamine stretch. At the expansion of polyQ above 39 residues, it has an inherent propensity to form amyloid-like fibrils and aggregates, and is responsible for Huntington's disease. The results of the experiments show that expression of the eGFP in fusion with the 97Q protein leads to the decrease of the eGFP fluorescence lifetime by similar to 300 ps. This phenomenon does not appear in Hsp104-deficient cells, where the aggregation in polyQ is prevented. We demonstrate that the lifetime decrease observed is related to the aggregation per se and discuss the possible role of refractive index and homo-FRET in these dynamics. (C) 2010 Society of Photo-Optical Instrumentation Engineers. [DOI: 10.1117/1.3290821]