Ferguson Plot Analysis of Chaperone ClpB from Moderate Halophile.

Teruo Akuta, Yui Tomioka, Tomoto Ura, Masataka Nakagawa, Tsutomu Arakawa
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Abstract

The Ferguson plot is a simple method for determining the molecular weight of native proteins and their complexes. In this study, we tested the validity of the Ferguson plot based on agarose native gel electrophoresis using multimeric chaperone protein, ClpB, derived from a moderate halophile that forms a native hexamer. The Ferguson plot showed a single band with a molecular weight of 1,500 kDa, approximately twice the size of the native hexamer. This result is consistent with the structure of other chaperons that form a double ring assembly comprising two hexameric units, i.e., a dodecamer. Supporting this, dynamic light scattering experiment showed two peaks, which likely correspond to the hexamer and dodecamer structures.

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中等亲盐菌伴生物ClpB的Ferguson图分析。
弗格森图是测定天然蛋白及其复合物分子量的一种简单方法。在这项研究中,我们使用多聚体伴侣蛋白ClpB测试了基于琼脂糖天然凝胶电泳的Ferguson图的有效性,ClpB来源于一种形成天然六聚体的中等亲盐菌。Ferguson图显示了一个分子量为1500 kDa的单条带,大约是天然六聚体的两倍。这一结果与其他伴子的结构一致,它们形成由两个六聚体单元组成的双环组装,即十二聚体。动态光散射实验显示出两个峰,可能对应于六聚体和十二聚体结构。
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