Development of glutathione-coupled cantilever for the single-molecule force measurement by scanning force microscopy
Development of glutathione-coupled cantilever for the single-molecule force measurement by scanning force microscopy
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DOI：
10.1016/j.febslet.2006.06.032
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发表时间：
2006-07-10
期刊：
影响因子：
3.5
通讯作者：
Takeyasu, Kunio
中科院分区：
文献类型：
--
作者：
Yoshimura, Shige H.;Takahashi, Hirohide;Takeyasu, Kunio
单分子测力的准确性和保真度在很大程度上取决于感兴趣的分子如何附着到固体衬底表面(珠子、悬臂、盖子玻璃等)。在不影响蛋白质结构和酶功能的情况下，蛋白质的定点附着一直是人们主要关注的问题。在这里，我们建立了一个谷胱甘肽偶联的悬臂，任何谷胱甘肽S转移酶(GST)的融合蛋白都可以以期望的方向附着在上面。谷胱甘肽和谷胱甘肽之间的破碎力平均接近100pN。利用这个悬臂梁，我们成功地测量了Importinα和Importinβ之间的相互作用力。(C)2006年，由Elsevier B.V.代表欧洲生化学会联合会出版。
The accuracy and the fidelity of a single-molecule force measurement largely rely on how the molecule of interest is attached to the solid substrate surface (bead, cantilever, cover glass and etc.). A site-specific attachment of a protein without affecting its structure and enzymatic function has been a major concern. Here, we established a glutathione-coupled cantilever to which any glutathione S-transferase (GST)-fused proteins can be attached in a desired direction. The rupture force between glutathione and GST was similar to 100 pN on average. By using this cantilever, we succeeded in measuring the interaction force between importin alpha and importin beta. (c) 2006 Published by Elsevier B.V. on behalf of the Federation of European Biochemical Societies.