Protein analysis by dynamic light scattering: Methods and techniques for students

Authors

  • Bernard Lorber,

    Corresponding author
    1. Research team Traduction mitochondriale et pathologies, Architecture et Réactivité de l'ARN, Université de Strasbourg, CNRS, IBMC, 15 rue René Descartes, F-67084 Strasbourg, France
    • Research team Traduction mitochondriale et pathologies, Architecture et Réactivité de l'ARN, Université de Strasbourg, CNRS, IBMC, 15 rue René Descartes, F-67084 Strasbourg, France
    Search for more papers by this author
    • Frédéric Fischer,

      1. Research team Fonctions et dynamiques des nanomachines de l'appareil de traduction, Architecture et Réactivité de l'ARN, Université de Strasbourg, CNRS, IBMC, 15 rue René Descartes, F-67084 Strasbourg, France
      Search for more papers by this author
      • F. Fischer and B. Lorber contributed equally to this work.

    • Marc Bailly,

      1. Research team Fonctions et dynamiques des nanomachines de l'appareil de traduction, Architecture et Réactivité de l'ARN, Université de Strasbourg, CNRS, IBMC, 15 rue René Descartes, F-67084 Strasbourg, France
      Current affiliation:
      1. Department of Microbiology and Cell Science, P.O. Box 110700, University of Florida, Gainesville, FL 32611-0700, USA
      Search for more papers by this author
    • Hervé Roy,

      1. Research team Fonctions et dynamiques des nanomachines de l'appareil de traduction, Architecture et Réactivité de l'ARN, Université de Strasbourg, CNRS, IBMC, 15 rue René Descartes, F-67084 Strasbourg, France
      Current affiliation:
      1. Burnett School of Biomedical Sciences, University of Central Florida, 12722 Research Parkway, Orlando, FL 32826, USA
      Search for more papers by this author
    • Daniel Kern

      1. Research team Fonctions et dynamiques des nanomachines de l'appareil de traduction, Architecture et Réactivité de l'ARN, Université de Strasbourg, CNRS, IBMC, 15 rue René Descartes, F-67084 Strasbourg, France
      Search for more papers by this author

    • This work is supported by the Université de Strasbourg, the French Ministère de l'Enseignement Supérieur et de la Recherche, the Association pour la Recherche sur le Cancer (ARC), the Centre National de la Recherche Scientifique (CNRS), French National Program Investissement d'Avenir (Labex Mitocross).

    Abstract

    Dynamic light scattering (DLS) analyses are routinely used in biology laboratories to detect aggregates in macromolecular solutions, to determine the size of proteins, nucleic acids, and complexes or to monitor the binding of ligands. This article is written for graduate and undergraduate students with access to DLS and for faculty members who wish to incorporate DLS into a lab activity, a practical course or research. It reviews the basic concepts of light scattering measurements and addresses four critical aspects of the analysis and interpretation of DLS results. To ensure reproducible quantitative data, attention should be paid to controlling the preparation and handling of proteins or assemblies because variations in the state of aggregation, induced by minor changes in experimental condition or technique, might compromise DLS results and affect protein activity. Variables like temperature, solvent viscosity, and inter-particle interactions may also influence particle size determination. Every point is illustrated by case studies, including a commercially available albumin, a small RNA virus isolated from plants, as well as four soluble proteins and a ribonucleoprotein assembly purified and characterized by students in the frame of their master degree. © 2012 by The International Union of Biochemistry and Molecular Biology

    Ancillary