Technical improvement to 2D-PAGE of rice organelle membrane proteins.
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.
Mikami
Satoshi
S
Laboratories of Molecular Life Science, Graduate School of Science and Technology, Niigata University, Japan.
Kishimoto
Tadashi
T
Hori
Hidetaka
H
Mitsui
Toshiaki
T
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*
D008565:Membrane Proteins; D010940:Plant Proteins