Controlled unzipping of a bacterial surface layer with atomic force microscopy
Controlled unzipping of a bacterial surface layer with atomic force microscopy
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DOI：
10.1073/pnas.96.23.13170
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发表时间：
1999-11-09
影响因子：
11.1
通讯作者：
Engel, A
中科院分区：
文献类型：
--
作者：
Müller, DJ;Baumeister, W;Engel, A
我们将高分辨率原子力显微镜（AFM）成像和力光谱相结合，以深入了解耐辐射球菌六边形填充中间层（HPI）中单个原聚物之间的相互作用力。在HPI层成像后，AFM手写笔通过强制手写笔与样品接触连接到单个原聚物上，以便进行力光谱实验。记录力延伸曲线后的HPI层成像允许粘附力与结构变化相关。通过使用这种方法，发现HPI层的单个原形成物在大约300 pN的拉力下被去除。此外，有可能依次解压缩由6个HPI原形成物形成的整个细菌孔隙。单个蛋白质的高分辨率AFM成像与分子内力的测定相结合，是在单分子水平上研究超分子结构机械稳定性的一种方法。
We have combined high-resolution atomic force microscopy (AFM) imaging and force spectroscopy to gain insight into the interaction forces between the individual protomers of the hexagonally packed intermediate (HPI) layer of Deinococcus radiodurans, After imaging the HPI layer, the AFM stylus was attached to individual protomers by enforced stylus-sample contact to allow force spectroscopy experiments. Imaging of the HPI layer after recording force-extension curves allowed adhesion forces to be correlated with structural alterations. By using this approach, individual protomers of the HPI layer were found to be removed at pulling forces of approximate to 300 pN. Furthermore, it was possible to sequentially unzip entire bacterial pores formed by six HPI protomers, The combination of high-resolution AFM imaging of individual proteins with the determination of their intramolecular forces is a method of studying the mechanical stability of supramolecular structures at the level of single molecules.