Two-dimensional gel electrophoresis of rat liver microsomal membrane proteins.
Total rat liver microsomal proteins are not suitable for isoelectric focusing in polyacrylamide gels, even in the presence of sodium dodecyl sulphate and excess non-ionic detergent; considerable quantities of protein form an aggregate in the isoelectric focusing gel. This prevents resolution of microsomal proteins by the increasingly popular two-dimensional electrophoresis technique employing isoelectric focusing followed by sodium dodecyl sulphate-polyacrylamide gel electrophoresis. The problem is caused by the extreme insolubility of some microsomal proteins, especially cytochrome P-450 species, which precipitate during isoelectric focusing. A selective extraction of microsomes with sodium deoxycholate excludes these poorly soluble proteins. The extracted proteins can then be resolved without difficulty by isoelectric focusing, and give excellent two-dimensional gel patterns showing more than 100 proteins, mainly in the pI range 5--7. The technique should be useful in studies on microsome protein topology and on changes in microsome composition.
D000818:Animals; D003577:Cytochrome P-450 Enzyme System / Q000032:analysis; D004591:Electrophoresis, Polyacrylamide Gel; D007425:Intracellular Membranes / Q000032:analysis*; D008297:Male; D008565:Membrane Proteins / Q000032:analysis*; D008862:Microsomes, Liver / Q000032:analysis*; D008970:Molecular Weight; D051381:Rats; D012967:Sodium Dodecyl Sulfate
D008565:Membrane Proteins; D012967:Sodium Dodecyl Sulfate; D003577:Cytochrome P-450 Enzyme System