Solubilization of the cytoplasmic membrane of Escherichia coli by Triton X-100.
Treatment of a partially purified preparation of cell walls of Escherichia coli with Triton X-100 at 23 C resulted in a solubilization of 15 to 25% of the protein. Examination of the Triton-insoluble material by electron microscopy indicated that the characteristic morphology of the cell wall was not affected by the Triton extraction. Contaminating fragments of the cytoplasmic membrane were removed by Triton X-100, including the fragments of the cytoplasmic membrane which were normally observed attached to the cell wall. Treatment of a partially purified cytoplasmic membrane fraction with Triton X-100 resulted in the solubilization of 60 to 80% of the protein of this fraction. Comparison of the Triton-soluble and Triton-insoluble proteins from the cell wall and cytoplasmic membrane fractions by polyacrylamide gel electrophoresis after removal of the Triton by gel filtration in acidified dimethyl formamide indicated that the detergent specifically solubilized proteins of the cytoplasmic membrane. The proteins solubilized from the cell wall fraction were qualitatively identical to those solubilized from the cytoplasmic membrane fraction, but were present in different proportions, suggesting that the fragments of cytoplasmic membrane which are attached to the cell wall are different in composition from the remainder of the cytoplasmic membrane of the cell. Treatment of unfractionated envelope preparations with Triton X-100 resulted in the solubilization of 40% of the protein, and only proteins of the cytoplasmic membrane were solubilized. Extraction with Triton thus provides a rapid and specific means of separating the proteins of the cell wall and cytoplasmic membrane of E. coli.
D001426:Bacterial Proteins / Q000032:analysis; D002021:Buffers; D002458:Cell Fractionation; D002462:Cell Membrane / Q000032:analysis / Q000187:drug effects*; D002473:Cell Wall / Q000032:analysis / Q000187:drug effects; D002499:Centrifugation, Density Gradient; D002850:Chromatography, Gel; D004126:Dimethylformamide; D004589:Electrophoresis, Disc; D004926:Escherichia coli / Q000032:analysis / Q000166:cytology* / Q000187:drug effects; D007930:Leucine; D008274:Magnesium; D008854:Microscopy, Electron; D012995:Solubility; D012997:Solvents / Q000494:pharmacology*; D013395:Sucrose; D013501:Surface-Active Agents / Q000494:pharmacology*; D014316:Tritium; D014443:Tyrosine
D001426:Bacterial Proteins; D002021:Buffers; D012997:Solvents; D013501:Surface-Active Agents; D014316:Tritium; D014443:Tyrosine; D013395:Sucrose; D004126:Dimethylformamide; D007930:Leucine; D008274:Magnesium