Sorbonne Universite

5 articles published in JoVE

• Bioengineering

  
  Reconstitution of Actin-Based Motility with Commercially Available Proteins Cécile Sykes¹, Julie Plastino^{1 1}Laboratoire de physique de l’Ecole Normale Supérieure, ENS, Université PSL, CNRS, Sorbonne Université, Université Paris Cité This protocol describes how to produce actin comets on the surfaces of beads using commercially available protein ingredients. Such systems mimic the protrusive structures found in cells, and can be used to examine physiological mechanisms of force production in a simplified way.

• 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 Beber^1,7, Aurélie Di Cicco¹, Bassam Hajj¹, François Iv², Manos Mavrakis², Gijsje H. Koenderink³, João T. Cabral⁴, Michaël Trichet⁵, Stéphanie Mangenot*⁶, Aurélie Bertin*^{1 1}Laboratoire 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. Koenderink^{1 1}Department 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.

• Chemistry

  
  Thermochemical Studies of Ni(II) and Zn(II) Ternary Complexes Using Ion Mobility-Mass Spectrometry Anna J. Corrales¹, Anna V. Arredondo¹, Amber A. Flores¹, Chloe L. Duvak¹, Charles L. Mitchell¹, Riccardo Spezia², Laurence A. Angel^{1 1}Department of Chemistry, Texas A&M University-Commerce, ²Laboratoire de Chimie Théorique, Sorbonne Université This article describes an experimental protocol using electrospray-ion mobility-mass spectrometry, semi-empirical quantum calculations, and energy-resolved threshold collision-induced dissociation to measure the relative thermochemistry of the dissociation of related ternary metal complexes.

• Biology

  
  Analyzing Oxidative Stress in Murine Intestinal Organoids using Reactive Oxygen Species-Sensitive Fluorogenic Probe Aline Stedman^1,3, Antonin Levy^1,4, Philippe J. Sansonetti^1,2,5, Giulia Nigro^{1,6 1}Molecular Microbial Pathogenesis Unit, Institut Pasteur, ²Chaire de Microbiologie et Maladies Infectieuses, Collège de France, ³Institut de Biologie Paris Seine (IBPS) - Developmental Biology Unit, Sorbonne Université, CNRS UMR7622, INSERM U1156, ⁴Molecular Radiotherapy, INSERM U1030, Gustave Roussy, Université Paris-Saclay, ⁵The Center for Microbes, Development and Health, Institut Pasteur Shanghai and Chinese Academy of Sciences, ⁶Microenvironment and Immunity Unit, Institut Pasteur The present protocol describes a method to detect reactive oxygen species (ROS) in the intestinal murine organoids using qualitative imaging and quantitative cytometry assays. This work can be potentially extended to other fluorescent probes to test the effect of selected compounds on ROS.