Kroeck Kyle G, Qiu Weihua, Catalano Claudio, Trinh Thi Kim Hoang, Guo Youzhong
Department of Medicinal Chemistry, School of Pharmacy, Virginia Commonwealth University; Institute for Structural Biology, Drug Discovery and Development, School of Pharmacy, Virginia Commonwealth University.
Department of Medicinal Chemistry, School of Pharmacy, Virginia Commonwealth University; Institute for Structural Biology, Drug Discovery and Development, School of Pharmacy, Virginia Commonwealth University;
J Vis Exp. 2020 Jul 16(161). doi: 10.3791/61298.
Protein-protein interactions in cell membrane systems play crucial roles in a wide range of biological processes- from cell-to-cell interactions to signal transduction; from sensing environmental signals to biological response; from metabolic regulation to developmental control. Accurate structural information of protein-protein interactions is crucial for understanding the molecular mechanisms of membrane protein complexes and for the design of highly specific molecules to modulate these proteins. Many in vivo and in vitro approaches have been developed for the detection and analysis of protein-protein interactions. Among them the structural biology approach is unique in that it can provide direct structural information of protein-protein interactions at the atomic level. However, current membrane protein structural biology is still largely limited to detergent-based methods. The major drawback of detergent-based methods is that they often dissociate or denature membrane protein complexes once their native lipid bilayer environment is removed by detergent molecules. We have been developing a native cell membrane nanoparticle system for membrane protein structural biology. Here, we demonstrate the use of this system in the analysis of protein-protein interactions on the cell membrane with a case study of the oligomeric state of AcrB.
细胞膜系统中的蛋白质-蛋白质相互作用在广泛的生物过程中发挥着关键作用——从细胞间相互作用到信号转导；从感知环境信号到生物反应；从代谢调节到发育控制。蛋白质-蛋白质相互作用的准确结构信息对于理解膜蛋白复合物的分子机制以及设计高度特异性分子来调节这些蛋白质至关重要。已经开发了许多体内和体外方法用于检测和分析蛋白质-蛋白质相互作用。其中，结构生物学方法具有独特之处，因为它可以在原子水平上提供蛋白质-蛋白质相互作用的直接结构信息。然而，当前的膜蛋白结构生物学在很大程度上仍然局限于基于去污剂的方法。基于去污剂的方法的主要缺点是，一旦去污剂分子去除了其天然脂质双层环境，它们通常会使膜蛋白复合物解离或变性。我们一直在为膜蛋白结构生物学开发一种天然细胞膜纳米颗粒系统。在这里，我们通过AcrB寡聚状态的案例研究展示了该系统在分析细胞膜上蛋白质-蛋白质相互作用中的应用。
