Static light scattering to characterize membrane proteins in detergent solution.
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.
Slotboom
Dirk Jan
DJ
University of Groningen, Groningen Biomolecular Science and Biotechnology Institute, Nijenborgh 4, 9747 AG Groningen, The Netherlands. d.j.slotboom@rug.nl
Duurkens
Ria H
RH
Olieman
Kees
K
Erkens
Guus B
GB
D002499:Centrifugation, Density Gradient; D002850:Chromatography, Gel / Q000379:methods; D003902:Detergents; D008027:Light; D008433:Mathematics; D008565:Membrane Proteins / Q000737:chemistry*; D008823:Micelles; D012542:Scattering, Radiation; D012996:Solutions
D003902:Detergents; D008565:Membrane Proteins; D008823:Micelles; D012996:Solutions