title: Separation of membrane protein complexes by native LDS-PAGE.
abstract: Gel electrophoresis has become one of the most important methods for the analysis of proteins and protein complexes in a molecular weight range of 1-10(7) kDa. The separation of membrane protein complexes remained challenging to standardize until the demonstration of Blue Native PAGE in 1991 [1] and Clear Native PAGE in 1994 [2]. We present a robust protocol for high-resolution separation of photosynthetic complexes from Arabidopsis thaliana using lithium dodecyl sulfate as anion in a modified Blue Native PAGE (LDS-PAGE). Here, non-covalently bound chlorophyll is used as a sensitive probe to characterize the assembly/biogenesis of the pigment-protein complexes essential for photosynthesis. The high fluorescence yield recorded from chlorophyll-binding protein complexes can also be used to establish the separation of native protein complexes as an electrophoretic standard.
authors.affiliation: Center for Organelle Research (CORE), University of Stavanger, Stavanger, Norway.
authors.forename: Janine
authors.initials: J
authors.lastname: Arnold
authors.forename: Alexey
authors.initials: A
authors.lastname: Shapiguzov
authors.forename: Geoffrey
authors.initials: G
authors.lastname: Fucile
authors.forename: Jean-David
authors.initials: JD
authors.lastname: Rochaix
authors.forename: Michel
authors.initials: M
authors.lastname: Goldschmidt-Clermont
authors.forename: Lutz Andreas
authors.initials: LA
authors.lastname: Eichacker
mesh_terms: D017360:Arabidopsis / Q000378:metabolism*; D029681:Arabidopsis Proteins / Q000302:isolation & purification*; D004591:Electrophoresis, Polyacrylamide Gel / Q000379:methods*; D008565:Membrane Proteins / Q000302:isolation & purification*; D012967:Sodium Dodecyl Sulfate / Q000737:chemistry*; D012995:Solubility; D020524:Thylakoids / Q000378:metabolism
chemical_list: D029681:Arabidopsis Proteins; D008565:Membrane Proteins; D012967:Sodium Dodecyl Sulfate; C028913:dodecyl sulfate