.png){kind=icon}
返回
Single-Molecule Manipulation in Zero-Mode Waveguides
.png){kind=icon}
OA
AI
DOI：10.1002/smll.201906740.png)
.png){kind=icon}
原文求助.png){kind=icon}
分享.png){kind=icon}
收藏
摘要
En 中文
The mechanobiology of receptor-ligand interactions and force-induced enzymatic turnover can be revealed by simultaneous measurements of force response and fluorescence. Investigations at physiologically relevant high labeled substrate concentrations require total internal reflection fluorescence microscopy or zero mode waveguides (ZMWs), which are difficult to combine with atomic force microscopy (AFM). A fully automatized workflow is established to manipulate single molecules inside ZMWs autonomously with noninvasive cantilever tip localization. A protein model system comprising a receptor-ligand pair of streptavidin blocked with a biotin-tagged ligand is introduced. The ligand is pulled out of streptavidin by an AFM cantilever leaving the receptor vacant for reoccupation by freely diffusing fluorescently labeled biotin, which can be detected in single-molecule fluorescence concurrently to study rebinding rates. This work illustrates the potential of the seamless fusion of these two powerful single-molecule techniques.
Keyword:
force activation
mechanosensing
single-molecule fluorescence
zero mode waveguides
期刊
IF:
12.1
论文数:
3.0W
被引数:
16.4W
机构
引用论文
Genetically encoded short peptide tag for versatile protein labeling by Sfp phosphopantetheinyl transferase遗传编码的短肽标签，用于通过Sfp磷酸泛酰巯基乙胺基转移酶进行通用蛋白质标记
OAAI
OAAI分享收藏分享收藏分享收藏A streptavidin variant with slower biotin dissociation and increased mechanostability.png)
NATURE METHODSOAAI
NATURE METHODSIF32.1
OAAI分享收藏
查看更多内容