title: Specific removal of proteins from the envelope of Escherichia coli by protease treatments.
abstract: When the envelope fraction of Escherichia coli was treated by trypsin, about 40% of total envelope proteins were removed from the fraction without changing its phospholipid content. Analysis of envelope proteins by acrylamide gel electrophoresis in 0.5% sodium dodecyl sulfate revealed that trypsin treatment was very specific; one of the major proteins (molecular weight, 38,000) and all proteins of molecular weight greater than 70,000 were completely removed by the treatment. On the other hand, three other major proteins were found to be resistant to the treatment, including protein Y, which was previously shown to be related to deoxyribonucleic acid replication. The trypsin treatment of the envelope fractions composed of a five electron-dense layered structure formed vesicles with a triple-layered membrane (two electron-dense layers). Pronase treatment of the envelope fraction removed about 60% of the envelope proteins without changing its phospholipid content. A major protein of molecular weight of 58,000 was found to be the only protein resistant to the Pronase treatment. Application of these treatments is useful for purification and structural studies of envelope proteins.
mesh_terms: D000596:Amino Acids; D001426:Bacterial Proteins / Q000032:analysis / Q000302:isolation & purification*; D002247:Carbon Isotopes; D002458:Cell Fractionation; D002473:Cell Wall / Q000032:analysis*; D004591:Electrophoresis, Polyacrylamide Gel; D004926:Escherichia coli / Q000032:analysis* / Q000166:cytology; D005944:Glucosamine; D008970:Molecular Weight; D010759:Phosphorus Isotopes; D011402:Pronase / Q000378:metabolism*; D012967:Sodium Dodecyl Sulfate; D013194:Staining and Labeling; D014316:Tritium; D014357:Trypsin
chemical_list: D000596:Amino Acids; D001426:Bacterial Proteins; D002247:Carbon Isotopes; D010759:Phosphorus Isotopes; D014316:Tritium; D012967:Sodium Dodecyl Sulfate; D014357:Trypsin; D011402:Pronase; D005944:Glucosamine