Matos Anna Livia L, Grill David, Kudruk Sergej, Heitzig Nicole, Galla Hans-Joachim, Gerke Volker, Rescher Ursula
Institute of Medical Biochemistry, Center for Molecular Biology of Inflammation, University of Münster; Cluster of Excellence 'Cells in Motion', University of Münster.
Institute of Biochemistry, University of Münster; Cluster of Excellence 'Cells in Motion', University of Münster.
J Vis Exp. 2018 Nov 1(141). doi: 10.3791/58224.
The dissipative quartz crystal microbalance technique is a simple and label-free approach to measure simultaneously the mass uptake and viscoelastic properties of the absorbed/immobilized mass on sensor surfaces, allowing the measurements of the interaction of proteins with solid-supported surfaces, such as lipid bilayers, in real-time and with a high sensitivity. Annexins are a highly conserved group of phospholipid-binding proteins that interact reversibly with the negatively charged headgroups via the coordination of calcium ions. Here, we describe a protocol that was employed to quantitatively analyze the binding of annexin A2 (AnxA2) to planar lipid bilayers prepared on the surface of a quartz sensor. This protocol is optimized to obtain robust and reproducible data and includes a detailed step-by-step description. The method can be applied to other membrane-binding proteins and bilayer compositions.
耗散石英晶体微天平技术是一种简单且无需标记的方法，可同时测量传感器表面吸收/固定物质的质量摄取和粘弹性特性，能够实时且高灵敏度地测量蛋白质与固体支持表面（如脂质双层）之间的相互作用。膜联蛋白是一组高度保守的磷脂结合蛋白，通过钙离子的配位作用与带负电荷的头部基团可逆地相互作用。在此，我们描述了一种用于定量分析膜联蛋白A2（AnxA2）与在石英传感器表面制备的平面脂质双层结合的方案。该方案经过优化以获得可靠且可重复的数据，并包含详细的逐步说明。该方法可应用于其他膜结合蛋白和双层组成。
