de Franceschi Nicola, Alqabandi Maryam, Weissenhorn Winfried, Bassereau Patricia
Laboratoire Physico Chimie Curie, Institut Curie, PSL Research University, CNRS UMR168, Paris 75005, France.
Sorbonne Universite, Paris 75005, France.
Bio Protoc. 2019 Jul 5;9(13):e3294. doi: 10.21769/BioProtoc.3294.
investigation of the interaction between proteins and positively curved membranes can be performed using a classic nanotube pulling method. However, characterizing protein interaction with negatively curved membranes still represents a formidable challenge. Here, we describe our recently developed approach based on laser-triggered Giant Unilamellar Vesicles (GUVs) fusion. Our protocol allows sequential addition of proteins to a negatively curved membrane, while at the same time controlling the buffer composition, lipid composition and membrane tension. Moreover, this method does not require a step of protein detachment, greatly simplifying the process of protein encapsulation over existing methods.
蛋白质与正弯曲膜之间相互作用的研究可以使用经典的纳米管拉伸方法来进行。然而，表征蛋白质与负弯曲膜的相互作用仍然是一项艰巨的挑战。在这里，我们描述了我们最近开发的基于激光触发的巨型单层囊泡（GUVs）融合的方法。我们的方案允许将蛋白质顺序添加到负弯曲膜中，同时控制缓冲液组成、脂质组成和膜张力。此外，这种方法不需要蛋白质分离步骤，与现有方法相比大大简化了蛋白质封装过程。
