title: Tetra detector analysis of membrane proteins.
abstract: Well-characterized membrane protein detergent complexes (PDC) that are pure, homogenous, and stable, with minimized excess detergent micelles, are essential for functional assays and crystallization studies. Procedural steps to measure the mass, size, shape, homogeneity, and molecular composition of PDCs and their host detergent micelles using size-exclusion chromatography (SEC) with a Viscotek Tetra Detector Array (TDA; absorbance, refractive index, light scattering, and viscosity detectors) are presented in this unit. The value of starting with a quality PDC sample, the precision and accuracy of the results, and the use of a digital benchtop refractometer are emphasized. An alternate and simplified purification and characterization approach using SEC with dual absorbance and refractive index detectors to optimize detergent and lipid concentration while measuring the PDC homogeneity is also described. Applications relative to purification and characterization goals are illustrated as well.
authors.affiliation: Department of Biochemistry and Biophysics, University of California San Francisco, San Francisco, California.
authors.forename: Larry J W
authors.initials: LJW
authors.lastname: Miercke
authors.affiliation: Department of Biochemistry and Biophysics, University of California San Francisco, San Francisco, California.
authors.forename: Rebecca A
authors.initials: RA
authors.lastname: Robbins
authors.affiliation: Department of Biochemistry and Biophysics, University of California San Francisco, San Francisco, California.
authors.forename: Robert M
authors.initials: RM
authors.lastname: Stroud
mesh_terms: D002850:Chromatography, Gel / Q000379:methods*; D003902:Detergents / Q000737:chemistry*; D008565:Membrane Proteins / Q000032:analysis*; D008823:Micelles; D012031:Refractometry / Q000379:methods*
chemical_list: D003902:Detergents; D008565:Membrane Proteins; D008823:Micelles
keywords: differential pressure viscometer; intrinsic viscosity; membrane proteins; refractive index; tetra detector array and analysis
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