Examination of the protein composition of the cell envelope of Escherichia coli by polyacrylamide gel electrophoresis.
An envelope preparation containing the cell wall and cytoplasmic membrane of Escherichia coli was obtained by breaking the cells with a French pressure cell and sedimentating the envelope fraction by ultracentrifugation. This fraction was prepared for polyacrylamide gel electrophoresis by dissolving the protein in an acidified N,N'-dimethylformamide, removing lipids by gel filtration in the same organic solvent and removing the solvent by dialysis against aqueous urea solutions. More than 80% of the total protein of the envelope fraction was recovered in soluble form. Electrophoresis on sodium dodecyl sulfate-containing gels yielded from 20 to 30 well-resolved bands of protein. One major protein band was observed on the gels. This protein had a molecular weight of 44,000 and accounted for as much as 40% of the total protein of the envelope fraction. A double-labeling technique was used to examine the protein composition of the envelope fraction from cells grown under different sets of conditions which result in large changes in the levels of membrane-bound oxidative enzymes. These changes in growth conditions resulted in only minor alterations in the protein profiles observed on the gels, suggesting that this organism is able to adapt to changes in growth environment with only minor modifications of the major proteins of the cell envelope.
D000179:Acrylates; D001426:Bacterial Proteins / Q000032:analysis*; D002247:Carbon Isotopes; D002462:Cell Membrane / Q000032:analysis; D002473:Cell Wall / Q000032:analysis; D002850:Chromatography, Gel; D003470:Culture Media; D003956:Dialysis; D004126:Dimethylformamide; D004586:Electrophoresis; D004926:Escherichia coli / Q000032:analysis* / Q000201:enzymology / Q000254:growth & development; D005782:Gels; D005947:Glucose / Q000378:metabolism; D005990:Glycerol / Q000378:metabolism; D007930:Leucine; D008970:Molecular Weight; D010088:Oxidoreductases / Q000032:analysis; D010100:Oxygen; D012964:Sodium; D013431:Sulfates; D014316:Tritium; D014443:Tyrosine; D014461:Ultracentrifugation
D000179:Acrylates; D001426:Bacterial Proteins; D002247:Carbon Isotopes; D003470:Culture Media; D005782:Gels; D013431:Sulfates; D014316:Tritium; D014443:Tyrosine; D004126:Dimethylformamide; D012964:Sodium; D010088:Oxidoreductases; D007930:Leucine; D005947:Glucose; D005990:Glycerol; D010100:Oxygen