Ultrasonication-dependent acceleration of amyloid fibril formation.
Ultrasonication-dependent acceleration of amyloid fibril formation.
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DOI：
10.1016/j.jmb.2011.07.069
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发表时间：
2011-09
影响因子：
5.6
通讯作者：
M. So;H. Yagi;K. Sakurai;H. Ogi;H. Naiki;Y. Goto
中科院分区：
文献类型：
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作者：
M. So;H. Yagi;K. Sakurai;H. Ogi;H. Naiki;Y. Goto
淀粉样原纤维，类似于晶体，通过成核和生长形成。由于成核的高自由能势垒，淀粉样原纤维的自发形成只有在很长的滞后期才会发生。超声有助于诱导淀粉样蛋白成核，从而形成原纤维，而使用含有硫黄素T荧光的微孔板读卡器适合同时检测许多样品中的原纤维。结合超声和微孔板阅读器的使用，我们提出了一种有效的方法来研究蛋白质形成淀粉样原纤维的潜力。β2-微球蛋白（一种导致透析相关淀粉样变性的淀粉样蛋白）在pH为2.5时几分钟内就形成了原纤维。即使在中性pH条件下，原纤维的形成也需要1.5 h的滞后时间。结果表明，原纤维的形成是一种物理反应，在很大程度上受到高自由能势垒的限制，超声波可以有效地减少这种反应。这种方法将有助于开发蛋白质淀粉样变性的高通量测定。
Amyloid fibrils, similar to crystals, form through nucleation and growth. Because of the high free-energy barrier of nucleation, the spontaneous formation of amyloid fibrils occurs only after a long lag phase. Ultrasonication is useful for inducing amyloid nucleation and thus for forming fibrils, while the use of a microplate reader with thioflavin T fluorescence is suitable for detecting fibrils in many samples simultaneously. Combining the use of ultrasonication and microplate reader, we propose an efficient approach to studying the potential of proteins to form amyloid fibrils. With β2-microglobulin, an amyloidogenic protein responsible for dialysis-related amyloidosis, fibrils formed within a few minutes at pH 2.5. Even under neutral pH conditions, fibrils formed after a lag time of 1.5 h. The results propose that fibril formation is a physical reaction that is largely limited by the high free-energy barrier, which can be effectively reduced by ultrasonication. This approach will be useful for developing a high-throughput assay of the amyloidogenicity of proteins.