Martín-González Natalia, Ortega-Esteban Alvaro, Moreno-Madrid F, Llauró Aida, Hernando-Pérez Mercedes, de Pablo Pedro J
Departamento de Física de la Materia Condensada, C-3, Universidad Autónoma de Madrid, Ctra. de Colmenar Viejo, Km 15, 28049, Madrid, Spain.
Department of Physiology & Biophysics, University of Washington, Seattle, WA, 98195, USA.
Methods Mol Biol. 2018;1665:281-296. doi: 10.1007/978-1-4939-7271-5_15.
In Atomic Force Microscopy (AFM) the probe is a nanometric tip located at the end of a microcantilever which palpates the specimen under study as a blind person uses a white cane. In this way AFM allows obtaining nanometric resolution images of individual protein shells, such as viruses, in liquid milieu. Beyond imaging, AFM also enables the manipulation of single protein cages, and the characterization a variety physicochemical properties able of inducing any measurable mechanical perturbation to the microcantilever that holds the tip. In this chapter we start revising some recipes for adsorbing protein shells on surfaces. Then we describe several AFM approaches to study individual protein cages, ranging from imaging to spectroscopic methodologies devoted to extracting physical information, such as mechanical and electrostatic properties.
在原子力显微镜(AFM)中,探针是位于微悬臂梁末端的纳米级尖端,它像盲人使用白手杖一样探测被研究的样本。通过这种方式,AFM能够在液体环境中获得单个蛋白质外壳(如病毒)的纳米级分辨率图像。除了成像,AFM还能够对单个蛋白质笼进行操作,并表征各种能够对固定尖端的微悬臂梁施加任何可测量机械扰动的物理化学性质。在本章中,我们首先回顾一些将蛋白质外壳吸附在表面的方法。然后,我们描述几种研究单个蛋白质笼的AFM方法,从成像到致力于提取物理信息(如机械和静电性质)的光谱方法。