Techniques for Monitoring Protein Misfolding and Aggregation in Vitro and in Living Cells.
Techniques for Monitoring Protein Misfolding and Aggregation in Vitro and in Living Cells.
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DOI：
10.1007/s11814-012-0060-x
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发表时间：
2012-06
期刊：
影响因子：
--
通讯作者：
Kwon I
中科院分区：
文献类型：
--
作者：
Gregoire S;Irwin J;Kwon I
蛋白质的错误折叠和聚集被认为是理解许多神经退行性疾病和重组生物药物生产的重要因素。因此，各种传统和现代技术已被用来监测蛋白质聚集在体外和活细胞。分别用分子探针、TEM/AFM和CD/FTIR分析技术对体外淀粉样蛋白原纤维的形成、形态和二级结构含量进行了监测。活细胞中的蛋白质聚集已经被基于荧光蛋白或酶的许多分子折叠报告分子定性或定量地监测。靶蛋白的聚集与融合至靶蛋白的折叠报告子的荧光或酶活性的变化直接相关，这允许非侵入性监测活细胞中靶蛋白的聚集。用于在体外和活细胞中监测蛋白质聚集的技术的进展极大地促进了对与神经退行性疾病相关的淀粉样蛋白聚集的分子机制的理解，优化培养条件以减少在活细胞中表达的生物药物的聚集，以及在寻找蛋白质聚集抑制剂中筛选小分子文库。
Protein misfolding and aggregation have been considered important in understanding many neurodegenerative diseases and recombinant biopharmaceutical production. Therefore, various traditional and modern techniques have been utilized to monitor protein aggregation in vitro and in living cells. Fibril formation, morphology and secondary structure content of amyloidogenic proteins in vitro have been monitored by molecular probes, TEM/AFM, and CD/FTIR analyses, respectively. Protein aggregation in living cells has been qualitatively or quantitatively monitored by numerous molecular folding reporters based on either fluorescent protein or enzyme. Aggregation of a target protein is directly correlated to the changes in fluorescence or enzyme activity of the folding reporter fused to the target protein, which allows non-invasive monitoring aggregation of the target protein in living cells. Advances in the techniques used to monitor protein aggregation in vitro and in living cells have greatly facilitated the understanding of the molecular mechanism of amyloidogenic protein aggregation associated with neurodegenerative diseases, optimizing culture conditions to reduce aggregation of biopharmaceuticals expressed in living cells, and screening of small molecule libraries in the search for protein aggregation inhibitors.
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DOI：
10.1016/j.bbapap.2010.04.001
发表时间：
2010-07
期刊：
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影响因子：
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作者：
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