Isolation of mycoplasma membranes by digitonin.
The cell membrane of Mycoplasma hominis was isolated by lysing the cells with digitonin. Electron microscopy and chemical, density gradient, and electrophoretic analyses of the membrane proteins showed the membranes so obtained, like those isolated by osmotic lysis, to be relatively free of cytoplasmic contaminants. Sensitivity to digitonin lysis depended on temperature but was not affected by Mg(2+) ions and was only slightly affected by the age of the culture. Accordingly, it seems that digitonin may be used for the isolation of cell membranes from sterol-requiring mycoplasmas that tend to be fairly resistant to osmotic lysis.
D001426:Bacterial Proteins / Q000032:analysis; D001431:Bacteriological Techniques; D001433:Bacteriolysis; D002458:Cell Fractionation; D002462:Cell Membrane / Q000032:analysis; D002784:Cholesterol / Q000378:metabolism; D003470:Culture Media; D003720:Densitometry; D004352:Drug Resistance, Microbial; D004589:Electrophoresis, Disc; D008274:Magnesium / Q000494:pharmacology; D008854:Microscopy, Electron; D009174:Mycoplasma / Q000166:cytology* / Q000254:growth & development / Q000378:metabolism; D009829:Oleic Acids / Q000378:metabolism; D009994:Osmolar Concentration; D010772:Phosphotungstic Acid; D012503:Saponins; D013194:Staining and Labeling; D013261:Sterols / Q000378:metabolism; D013696:Temperature; D014316:Tritium
D001426:Bacterial Proteins; D003470:Culture Media; D009829:Oleic Acids; D012503:Saponins; D013261:Sterols; D014316:Tritium; D010772:Phosphotungstic Acid; D002784:Cholesterol; D008274:Magnesium