title: Combined small angle X-ray solution scattering with atomic force microscopy for characterizing radiation damage on biological macromolecules.
abstract: Synchrotron radiation facilities are pillars of modern structural biology. Small-Angle X-ray scattering performed at synchrotron sources is often used to characterize the shape of biological macromolecules. A major challenge with high-energy X-ray beam on such macromolecules is the perturbation of sample due to radiation damage.
  
  
  
  By employing atomic force microscopy, another common technique to determine the shape of biological macromolecules when deposited on flat substrates, we present a protocol to evaluate and characterize consequences of radiation damage. It requires the acquisition of images of irradiated samples at the single molecule level in a timely manner while using minimal amounts of protein. The protocol has been tested on two different molecular systems: a large globular tetremeric enzyme (β-Amylase) and a rod-shape plant virus (tobacco mosaic virus). Radiation damage on the globular enzyme leads to an apparent increase in molecular sizes whereas the effect on the long virus is a breakage into smaller pieces resulting in a decrease of the average long-axis radius.
  
  
  
  These results show that radiation damage can appear in different forms and strongly support the need to check the effect of radiation damage at synchrotron sources using the presented protocol.
authors: ESRF, The European Synchrotron, 71 Avenue des Martyrs, Grenoble, 38000, France.|Present Address: CBS, Centre de Biochimie Structurale, CNRS UMR 5048-INSERM UMR 1054, 29, Rue de Navacelles, Montpellier, 34090, France.
authors: Luca
authors: L
authors: Costa
authors: ESRF, The European Synchrotron, 71 Avenue des Martyrs, Grenoble, 38000, France.|MIT, 77 Massachusetts Ave., Cambridge, 02139, MA, USA.
authors: Alexander
authors: A
authors: Andriatis
authors: ESRF, The European Synchrotron, 71 Avenue des Martyrs, Grenoble, 38000, France.
authors: Martha
authors: M
authors: Brennich
authors: Univ. Grenoble Alpes, 71 Avenue des Martyrs, Grenoble, 38044, France.|CNRS, IBS, 71 Avenue des Martyrs, Grenoble, 38044, France.|CEA, IBS, 71 Avenue des Martyrs, Grenoble, France.
authors: Jean-Marie
authors: JM
authors: Teulon
authors: Univ. Grenoble Alpes, 71 Avenue des Martyrs, Grenoble, 38044, France.|CNRS, IBS, 71 Avenue des Martyrs, Grenoble, 38044, France.|CEA, IBS, 71 Avenue des Martyrs, Grenoble, France.
authors: Shu-Wen W
authors: SW
authors: Chen
authors: Univ. Grenoble Alpes, 71 Avenue des Martyrs, Grenoble, 38044, France. jean-luc.pellequer@ibs.fr.|CNRS, IBS, 71 Avenue des Martyrs, Grenoble, 38044, France. jean-luc.pellequer@ibs.fr.|CEA, IBS, 71 Avenue des Martyrs, Grenoble, France. jean-luc.pellequer@ibs.fr.
authors: Jean-Luc
authors: 0000-0002-8944-2715
authors: JL
authors: Pellequer
authors: European Molecular Biology Laboratory, 71 Avenue des Martyrs, Grenoble, 38000, France.|Unit for Virus Host-Cell Interactions, Univ. Grenoble Alpes-EMBL-CNRS, 71 Avenue des Martyrs, Grenoble, 38000, France.|Faculty of Natural Sciences, Keele University, Keele, Staffordshire, UK.|Present Address: European XFEL GmbH, Holzkoppel 4, Schenefeld, 22869, Germany.
authors: Adam
authors: A
authors: Round
mesh_terms: D000681:Amylases / Q000737:chemistry* / Q000378:metabolism / Q000528:radiation effects; D027723:Ipomoea batatas / Q000201:enzymology; D018625:Microscopy, Atomic Force; D009532:Nickel / Q000737:chemistry; D020836:Protein Structure, Quaternary; D053838:Scattering, Small Angle; D014027:Tobacco Mosaic Virus / Q000737:chemistry / Q000528:radiation effects; D014961:X-Ray Diffraction; D014965:X-Rays
chemical_list: C022838:nickel chloride; D009532:Nickel; D000681:Amylases
keywords: Atomic force microscopy (AFM); Radiation damage; Small angle x-ray scattering (SAXS); Tobacco mosaic virus; β-Amylase