Department of Biochemistry and Molecular Biology, Complutense University, Madrid, Spain; Biochemistry, Faculty of Science and Engineering, Åbo Akademi University, Turku, Finland.
Department of Biochemistry and Molecular Biology, Complutense University, Madrid, Spain; Department of Biotechnology and Biomedicine, Technical University of Denmark, Kongens Lyngby, Denmark.
Methods Enzymol. 2021;649:307-339. doi: 10.1016/bs.mie.2020.12.017. Epub 2021 Jan 11.
Protein-lipid interactions are crucial events from a biochemical point of view, like the interaction of proteins with the cell plasma membrane, and their study is of great importance. Actinoporins are a very powerful tool to study this kind of interactions, since they are soluble proteins in an aqueous environment, capable of inserting into membranes when they have the adequate composition. In fact, actinoporins have been used to study protein-lipid interactions for many years now. Sometimes it is not possible to use real biological membranes in the experiments, so model membranes need to be used. This article aims to give a thorough description of many of the techniques used to study actinoporin-lipid interactions, using both biological and model membranes: Hemolysis, release of vesicles content, surface plasmon resonance, isothermal titration calorimetry, fluorescence-based measurements, etc. Some of these techniques measure the actinoporins activity and some measure their binding properties. The combination of all the techniques described can offer valuable information about the thermodynamics and the kinetics of the actinoporin-lipid interaction.
从生化角度来看，蛋白质-脂质相互作用是至关重要的事件，如蛋白质与细胞质膜的相互作用，因此对其进行研究具有重要意义。肌动蛋白是研究这种相互作用的一种非常有力的工具，因为它们在水相环境中是可溶性蛋白质，当具有适当的组成时能够插入到膜中。事实上，肌动蛋白已经被用于研究蛋白质-脂质相互作用多年。有时在实验中不可能使用真正的生物膜，因此需要使用模型膜。本文旨在详细描述使用生物和模型膜研究肌动蛋白-脂质相互作用的许多技术：溶血、囊泡内容物释放、表面等离子体共振、等温滴定量热法、荧光测量等。其中一些技术测量肌动蛋白的活性，而另一些技术则测量其结合特性。描述的所有技术的组合可以提供关于肌动蛋白-脂质相互作用热力学和动力学的有价值的信息。
