Static light scattering to characterize membrane proteins in detergent solution

Dirk Jan Slotboom*, Ria H. Duurkens, Kees Olieman, Guus B. Erkens

*Corresponding author for this work

Research output: Contribution to journalArticleAcademicpeer-review

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Abstract

Determination of the oligomeric state or the subunit stoichiometry of integral membrane proteins in detergent solution is notoriously difficult, because the amount of detergent (and lipid) associated with the proteins is usually not known. Only two classical methods (sedimentation equilibrium centrifugation and static light scattering) can measure directly the absolute molecular mass of a protein present in a protein/detergent micelle, without any assumption on the amount of detergent bound or the shape of, the proteins. Here the theoretical background and practical aspects of static light scattering analysis of membrane proteins are reviewed using a number of examples from our lab to highlight potential pitfalls. A brief comparison with sedimentation equilibrium centrifugation is given and a detailed protocol of how we perform light scattering analyses is provided. (C) 2008 Elsevier Inc. All rights reserved.

Original languageEnglish
Pages (from-to)73-82
Number of pages10
JournalMethods
Volume46
Issue number2
DOIs
Publication statusPublished - Oct-2008

Keywords

  • Membrane protein
  • Oligomeric state
  • Detergent
  • Molecular mass
  • Static light scattering
  • SIZE-EXCLUSION CHROMATOGRAPHY
  • REFRACTIVE-INDEX DETECTORS
  • BACILLUS-STEAROTHERMOPHILUS
  • TRANSPORTER
  • RECONSTITUTION
  • PURIFICATION
  • ABSORBENCY

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