Chapter 4. Analytical Ultracentrifugation: Membrane Protein Assemblies in the Presence of Detergent

  1. Prof. Eva Pebay-Peyroula PhD professor
  1. Christine Ebel1,
  2. Jesper V. Møller2 and
  3. Marc le Maire3

Published Online: 7 JAN 2008

DOI: 10.1002/9783527621224.ch4

Biophysical Analysis of Membrane Proteins: Investigating Structure and Function

Biophysical Analysis of Membrane Proteins: Investigating Structure and Function

How to Cite

Ebel, C., Møller, J. V. and Maire, M. l. (2007) Analytical Ultracentrifugation: Membrane Protein Assemblies in the Presence of Detergent, in Biophysical Analysis of Membrane Proteins: Investigating Structure and Function (ed E. Pebay-Peyroula), Wiley-VCH Verlag GmbH & Co. KGaA, Weinheim, Germany. doi: 10.1002/9783527621224.ch4

Editor Information

  1. Institut de Biologie Structurale Jean-Pierre Ebel, Université Joseph Fourier-CEA-CNRS, 41 rue Jules Horowitz, 38027 Grenoble cedex 1, France

Author Information

  1. 1

    CNRS, IBS, Laboratoire de Biophysique Moléculaire, 41 rue Jules Horowitz, 38027 Grenoble Cedex 1, France

  2. 2

    Institute of Physiology and Biophysics, University of Aarhus, Ole Worms Allé 1185, 8000 C, Aarhus C, Denmark

  3. 3

    CEA, iBiTecS, Service de Bioénergétique Biologie, Structurale et Mécanismes, Laboratoire Protéines membranaires, 91191 Gif sur Yvette Cedex, France

Publication History

  1. Published Online: 7 JAN 2008
  2. Published Print: 24 OCT 2007

ISBN Information

Print ISBN: 9783527316779

Online ISBN: 9783527621224

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Keywords:

  • biophysical analysis;
  • membrane proteins;
  • molecular interaction;
  • large assemblies;
  • analytical ultracentrifugation;
  • sedimentation equilibrium;
  • sedimentation velocity;
  • data analysis;
  • analytical ultracentrifugation;
  • SANS/SAXS

Summary

This chapter contains sections titled:

  • Introduction

  • Instrumentation and the Principle of Typical Experiments

  • General Theoretical Background

    • Equation of the Transport

    • The Macromolecular Parameters: RS, Mb, M, and Ṽ

    • The Svedberg Equation

      • Mean values of Mb and s

    • Non-Ideality

  • Membrane Proteins: Measurement of RS, Mb, M, and Ṽ

    • Composition and Molar Mass

    • Values of Ṽ

    • Buoyant Mass for Detergent-Solubilized Membrane Proteins, Mb*

    • Stokes Radius, Frictional Ratio

    • The Example of the Membrane Protein BmrA

  • Sedimentation Equilibrium Data Analysis

    • Equation of Sedimentation Equilibrium and Comments on the Experimental Set-Up

    • Simulation of Sedimentation Equilibrium for a Mixture of Particles

    • Analysis of Data

    • Matching of Surfactant and Solvent Densities

    • Determining the Association States and Dissociation Constant in the Presence of Non-Density-Matched Detergent

    • Dependency of Association Constants on Detergent Concentration

  • Sedimentation Velocity Data Analysis

    • Numerical Solutions of the Lamm Equation

    • Analysis in Terms of Non-Interacting Species: Principle

    • Analysis in Terms of Non-Interacting Species: Applications to Detergent and the Membrane Protein EmrE

    • c(s) Analysis: Principle

    • Sedimentation Velocity Simulation and c(s) Analysis for a Hypothetical Sample of Membrane Proteins

    • Example of Characterization of a Membrane Protein by Sedimentation Velocity

      • Association State of Na+-K+-ATPase Expressed in Pichia pastoris and of Sarcoplasmic Ca2+-ATPase

      • Complex Behavior in Solution of New Amphiphilic Compounds

      • The SH/SD Method

    • General Potentials of the c(s) Analysis per se as a Prelude to more Sophisticated Analysis

  • Analytical Ultracentrifugation and SANS/SAXS

  • Conclusions