Aix Marseille Univ, Centrale Marseille 2 articles published in JoVE Biology Bottom-Up In Vitro Methods to Assay the Ultrastructural Organization, Membrane Reshaping, and Curvature Sensitivity Behavior of Septins Brieuc Chauvin*1, Koyomi Nakazawa*1, Alexandre Beber1,7, Aurélie Di Cicco1, Bassam Hajj1, François Iv2, Manos Mavrakis2, Gijsje H. Koenderink3, João T. Cabral4, Michaël Trichet5, Stéphanie Mangenot*6, Aurélie Bertin*1 1Laboratoire Physico Chimie Curie, Institut Curie, PSL Research University, Sorbonne Université, 2Institut Fresnel, CNRS UMR7249, Aix Marseille Univ, Centrale Marseille, 3Department of Bionanoscience, Kavli Institute of Nanoscience Delft, Delft University of Technology, 4Department of Chemical Engineering, Imperial College London, 5Sorbonne Université, CNRS, Institut de Biologie Paris-Seine (IBPS), Service de microscopie électronique (IBPS-SME), 6Laboratoire Matière et Systèmes Complexes (MSC), Université Paris Cité, 7Institute of Biotechnology, Czech Academy of Sciences, BIOCEV Septins are cytoskeletal proteins. They interact with lipid membranes and can sense but also generate membrane curvature at the micron scale. We describe in this protocol bottom-up in vitro methodologies for analyzing membrane deformations, curvature-sensitive septin binding, and septin filament ultrastructure. Biochemistry Purification and Quality Control of Recombinant Septin Complexes for Cell-Free Reconstitution Gerard Castro-Linares1, Jeffrey den Haan1, Francois Iv2, Carla Silva Martins2, Aurélie Bertin3, Manos Mavrakis2, Gijsje H. Koenderink1 1Department of Bionanoscience, Kavli Institute of Nanoscience Delft, Delft University of Technology, 2Institut Fresnel, CNRS UMR7249, Aix Marseille Univ, Centrale Marseille, 3Institut Curie, Université PSL, Sorbonne Université In vitro reconstitution of cytoskeletal proteins is a vital tool to understand the basic functional properties of these proteins. The present paper describes a protocol to purify and assess the quality of recombinant septin complexes, which play a central role in cell division and migration.
