title: Technical improvement to 2D-PAGE of rice organelle membrane proteins.
abstract: Cytosolic and membrane-associated proteins prepared from rice cells were separated and compared by two different 2D-PAGE methods, isoelectric focusing (IEF)/SDS-PAGE and nonequilibrium pH gradient electrophoresis (NEPHGE)/SDS-PAGE. Although IEF/SDS-PAGE of the cytosolic proteins showed sufficient resolution, some mitochondrial and basic microsomal membrane-associated proteins were weakly or hardly detectable on the 2D gel. High-quality and -quantity separation of the organelle membrane-associated proteins was accomplished by NEPHGE/SDS-PAGE, the advantage of this method being more critical in tightly membrane-bound proteins that were unwashable with NaCl. These results indicate that NEPHGE/SDS-PAGE is a useful tool for the proteomic analysis of rice membrane-associated proteins.
authors.affiliation: Laboratories of Molecular Life Science, Graduate School of Science and Technology, Niigata University, Japan.
authors.forename: Satoshi
authors.initials: S
authors.lastname: Mikami
authors.forename: Tadashi
authors.initials: T
authors.lastname: Kishimoto
authors.forename: Hidetaka
authors.initials: H
authors.lastname: Hori
authors.forename: Toshiaki
authors.initials: T
authors.lastname: Mitsui
mesh_terms: D015180:Electrophoresis, Gel, Two-Dimensional / Q000379:methods*; D008565:Membrane Proteins / Q000302:isolation & purification*; D015388:Organelles / Q000737:chemistry*; D012275:Oryza / Q000737:chemistry*; D010940:Plant Proteins / Q000302:isolation & purification*
chemical_list: D008565:Membrane Proteins; D010940:Plant Proteins