Dynamic and Sequential Protein Reconstitution on Negatively Curved Membranes by Giant Vesicles Fusion
Nicola Franceschi,
Maryam Alqabandi,
Winfried Weissenhorn,
Patricia Bassereau
Affiliations
Nicola Franceschi
Laboratoire Physico Chimie Curie, Institut Curie, PSL Research University, CNRS UMR168, Paris 75005, FranceSorbonne Université, Paris 75005, France, Department of Bionanoscience, Kavli Institute of Nanoscience Delft, Delft University of Technology, Delft, the Netherlands
Maryam Alqabandi
Laboratoire Physico Chimie Curie, Institut Curie, PSL Research University, CNRS UMR168, Paris 75005, FranceSorbonne Université, Paris 75005, France
Winfried Weissenhorn
Institut de Biologie Structurale (IBS), Univ. Grenoble Alpes, CEA, CNRS, Grenoble 38000, France
Patricia Bassereau
Laboratoire Physico Chimie Curie, Institut Curie, PSL Research University, CNRS UMR168, Paris 75005, FranceSorbonne Université, Paris 75005, France
In vitro 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.
