Monitoring ligand-receptor interactions by photonic force microscopy.
Monitoring ligand-receptor interactions by photonic force microscopy.
复制标题
DOI：
10.1088/0957-4484/21/25/255102
复制
发表时间：
2010-06-25
期刊：
影响因子：
3.5
通讯作者：
Moy VT
中科院分区：
文献类型：
--
作者：
Jeney S;Mor F;Koszali R;Forró L;Moy VT
我们介绍了一种使用光子力显微镜(PFM)获取配体-受体相互作用的单分子力测量的方法。用光学陷阱操纵的生物素功能化小球和链霉亲和素功能化的盖片被用来测量不同拉力对单个链霉亲和素-生物素复合体寿命的影响。通过优化光学陷阱的设计和选择合适的磁珠尺寸，获得了超过50pN的拉力。基于附着的珠子三维(3D)热位置波动的幅度，我们能够选择由单个链霉亲和素-生物素复合体介导的珠子-盖片相互作用。此外，由于数据采集和分析的自动化，大大加快了实验系统的开发速度。在链霉亲和素和生物素之间的力依赖动力学测量中，我们观察到链霉亲和素-生物素复合体具有捕获键的性质，当拉力从10 pN增加到20pN时，寿命增加了10倍。我们还表明，硅珠比聚苯乙烯珠更适合作用力测量，因为可以从硅珠中提取出长度超过200 nm的聚苯乙烯系绳。
We introduce a method for the acquisition of single molecule force measurements of ligand —receptor interactions using the photonic force microscope (PFM). Biotin-functionalized beads, manipulated with an optical trap, and a streptavidin-functionalized coverslip were used to measure the effect of different pulling forces on the lifetime of individual streptavidin-biotin complexes. By optimizing the design of the optical trap and selection of the appropriate bead size, pulling forces in excess of 50 pN were achieved. Based on the amplitude of three dimensional (3D) thermal position fluctuations of the attached bead, we were able to select for a bead-coverslip interaction that was mediated by a single streptavidin-biotin complex. Moreover, the developed experimental system was greatly accelerated by automation of data acquisition and analysis. In force-dependent kinetic measurements carried out between streptavidin and biotin, we observed that the streptavidin-biotin complex exhibited properties of a catch bond with the lifetime increasing 10 fold when the pulling force increased from 10 to 20 pN. We also show that silica beads were more appropriate than polystyrene beads for the force measurements as polystyrene tethers, longer than 200 nm, could be extracted from the beads.
登录
查看更多内容
影响因子：
7.8
作者：
Pralle, A;Keller, P;Florin, E L;Simons, K;Horber, J K
通讯作者：
Horber, J K
DOI：
10.1083/jcb.200810002
发表时间：
2009-06-29
期刊：
The Journal of cell biology
影响因子：
--
作者：
Kong F;García AJ;Mould AP;Humphries MJ;Zhu C
通讯作者：
Zhu C
影响因子：
4
作者：
Guzman, Camilo;Flyvbjerg, Henrik;Jeney, Sylvia
通讯作者：
Jeney, Sylvia
影响因子：
2.9
作者：
Jeney, S;Stelzer, EHK;Florin, EL
通讯作者：
Florin, EL
影响因子：
8.6
作者：
Jeney, Sylvia;Lukic, Branimir;Forro, Laszlo
通讯作者：
Forro, Laszlo