Das Priyadip, Duanias-Assaf Tal, Reches Meital
Institute of Chemistry and The Center for Nanoscience and Nanotechnology, The Hebrew University of Jerusalem.
Institute of Chemistry and The Center for Nanoscience and Nanotechnology, The Hebrew University of Jerusalem;
J Vis Exp. 2017 Mar 6(121):54975. doi: 10.3791/54975.
The interactions between proteins or peptides and inorganic materials lead to several interesting processes. For example, combining proteins with minerals leads to the formation of composite materials with unique properties. In addition, the undesirable process of biofouling is initiated by the adsorption of biomolecules, mainly proteins, on surfaces. This organic layer is an adhesion layer for bacteria and allows them to interact with the surface. Understanding the fundamental forces that govern the interactions at the organic-inorganic interface is therefore important for many areas of research and could lead to the design of new materials for optical, mechanical and biomedical applications. This paper demonstrates a single-molecule force spectroscopy technique that utilizes an AFM to measure the adhesion force between either peptides or amino acids and well-defined inorganic surfaces. This technique involves a protocol for attaching the biomolecule to the AFM tip through a covalent flexible linker and single-molecule force spectroscopy measurements by atomic force microscope. In addition, an analysis of these measurements is included.
蛋白质或肽与无机材料之间的相互作用会引发几个有趣的过程。例如,将蛋白质与矿物质结合会形成具有独特性质的复合材料。此外,生物污垢这一不良过程是由生物分子(主要是蛋白质)在表面的吸附引发的。这个有机层是细菌的粘附层,使它们能够与表面相互作用。因此,了解支配有机 - 无机界面相互作用的基本力对于许多研究领域都很重要,并且可能会促成用于光学、机械和生物医学应用的新材料的设计。本文展示了一种单分子力谱技术,该技术利用原子力显微镜(AFM)来测量肽或氨基酸与明确的无机表面之间的粘附力。该技术涉及通过共价柔性接头将生物分子连接到AFM尖端的方案以及通过原子力显微镜进行的单分子力谱测量。此外,还包括对这些测量结果的分析。