Influence of protein immobilization on protein–protein interaction measured by scanning force spectroscopy
Influence of protein immobilization on protein–protein interaction measured by scanning force spectroscopy
复制标题
扫描力谱测量蛋白质固定对蛋白质-蛋白质相互作用的影响
DOI：
10.1002/pssa.201200680
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发表时间：
2013
期刊：
影响因子：
--
通讯作者：
C. Ziegler
中科院分区：
文献类型：
--
作者：
C. Rösch;C. Huber;C. Müller;N. Umanskaya;M. Hannig;C. Ziegler
为了更好地了解生物膜的形成，用扫描力光谱(SFS)研究了蛋白质在固定化蛋白质层上的粘附力。为此，将蛋白质层固定在表面的实验装置是重要的。考察了蛋白质固定化方法的影响，并分析了pH和接触时间的变化对蛋白质-蛋白质相互作用中粘附力测量的影响。以牛血清白蛋白(BSA)在涂覆牛血清白蛋白(BSA)的硅胶上的吸附为模型体系。采用自由吸附、戊二醛交联和硅烷化后共价蛋白质结合的方法对底物进行包覆。对于后者，比较了不同的硅烷沉积方法。根据固定化方法的不同，测得的蛋白质-蛋白质相互作用显示出明显的差异。蛋白质的共价吸附是获得稳定的、预吸附的蛋白质层用于SFS测量的一种很有前途的方法。涂覆牛血清白蛋白的悬臂梁在这些层上的粘附力似乎是由分子间的静电相互作用主导的。
For a better understanding of biofilm establishment, adhesion forces of proteins on immobilized protein layers were investigated by scanning force spectroscopy (SFS). For this purpose the experimental setup to immobilize protein layers on the surface is important. The influence of the method chosen to immobilize the proteins on the surface was examined and afterwards the effect of variations in pH and contact time on the measured adhesion forces in protein–protein interaction was analyzed. The adsorption of bovine serum albumin (BSA) on BSA‐coated silicon was used here as a model system. To coat the substrate, free adsorption, crosslinking with glutaraldehyde, and covalent protein binding to the surface after silanization were used. For the latter different deposition methods of the silane were compared. Depending on the immobilization method the measured protein–protein interactions show distinct differences. Covalent attachment of proteins is a promising approach to attain stable, pre‐adsorbed protein layers for SFS measurements. The adhesion forces of a BSA‐coated cantilever on such layers seem to be dominated by electrostatic interactions of the molecules.
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影响因子：
1.9
作者：
Hannig, M
通讯作者：
Hannig, M
影响因子：
6.7
作者：
Rüdiger, SG;Carlén, A;Olsson, J
通讯作者：
Olsson, J
DOI：
10.1016/j.colsurfa.2012.08.049
发表时间：
2012-11-20
影响因子：
5.2
作者：
Thakurta, Sanjukta Guha;Subramanian, Anuradha
通讯作者：
Subramanian, Anuradha
影响因子：
--
作者：
Li, J;Helmerhorst, EJ;Oppenheim, FG
通讯作者：
Oppenheim, FG
DOI：
--
发表时间：
1986
期刊：
影响因子：
--
作者：
J. Arends;J. Schuthof;J. Christoffersen
通讯作者：
J. Christoffersen