Atomic force microscopy captures length phenotypes in single proteins
Atomic force microscopy captures length phenotypes in single proteins
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DOI：
10.1073/pnas.96.20.11288
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发表时间：
1999-09-28
影响因子：
11.1
通讯作者：
Fernandez, JM
中科院分区：
文献类型：
--
作者：
Carrion-Vazquez, M;Marszalek, PE;Fernandez, JM
我们使用单蛋白质原子力显微镜技术来检测Ig模块中的长度表型。为了获得氨基酸分辨率，我们通过设计由多达12个相同重复组成的多蛋白来放大单个模块的机械特征。我们发现，在机械展开时，含有5个额外甘氨酸残基的突变多聚蛋白在模块的折叠核心上比野生型多聚蛋白每个模块延长20埃。相比之下，类似的插入在N或C末端附近不起作用。因此，我们的原子力显微镜测量很容易区分插入物的位置，并以类似于核磁共振和X射线结晶学的分辨率测量其大小。
We use single-protein atomic force microscopy techniques to detect length phenotypes in an Ig module. To gain amino acid resolution, we amplify the mechanical features of a single module by engineering polyproteins composed of up to 12 identical repeats. We show that on mechanical unfolding, mutant polyproteins containing five extra glycine residues added to the folded core of the module extend 20 Angstrom per module farther than the wild-type polyproteins. By contrast, similar insertions near the N or C termini have no effect. Hence, our atomic force microscopy measurements readily discriminate the location of the insert and measure its size with a resolution similar to that of NMR and x-ray crystallography.