Homogeneity of envelope proteins of Escherichia coli separated by gel electrophoresis in sodium dodecyl sulfate.
Some envelope proteins of Escherichia coli show variable behavior in acrylamide gel electrophoresis in 1% sodium dodecyl sulfate, depending upon the conditions of the solubilization. When solubilized in 1% sodium dodecyl sulfate at 70 C for 20 min, three distinct peaks (peaks 4, 6, and 7) are seen at molecular weights of 57,800, 44,300, and 38,400, respectively. However, when the envelope fractions are solubilized in 1% sodium dodecyl sulfate at 100 C for 5 min, or when they are treated with N, N-dimethylformamide at acidic pH before solubilization by our method, only a single peak at 48,000 molecular weight is observed in the molecular weight range mentioned above. That is, peaks 4 and 7 disappear and a new peak appears at the position overlapping with peak 6. Proteins isolated from peaks 4 and 7 show the similar molecular weight shifts to the new peak by the treatment at 100 C. No other peaks show any change by the heat treatment. The increase at the new peak is completely accounted for by the decrease at peaks 4 and 7, indicating that the new peak is composed of proteins from peaks 4, 6, and 7. However, it is concluded that these three peaks consist of distinctly different proteins for the following reasons: (i) they have different amino acid compositions, (ii) they show different solubilities in the nonionic detergent, Nonidet P-40, and as shown previously, (iii) peak 6 (protein Y) is related to deoxyribonucleic acid synthesis, and (iv) proteins in peaks 4, 6, and 7 have different resistance to proteolytic enzymes. Although the reasons for the anomalous molecular weight shifts of these peaks are not well understood at present, it is important to solubilize the E. coli envelope proteins by the standard method in order to investigate their properties and functions of the envelope proteins.
D000596:Amino Acids / Q000032:analysis; D001426:Bacterial Proteins / Q000032:analysis*; D002247:Carbon Isotopes; D002473:Cell Wall / Q000032:analysis*; D004126:Dimethylformamide; D004926:Escherichia coli / Q000032:analysis*; D006863:Hydrogen-Ion Concentration; D008722:Methods; D008970:Molecular Weight; D012967:Sodium Dodecyl Sulfate; D012995:Solubility; D013501:Surface-Active Agents; D013696:Temperature; D013997:Time Factors; D014316:Tritium
D000596:Amino Acids; D001426:Bacterial Proteins; D002247:Carbon Isotopes; D013501:Surface-Active Agents; D014316:Tritium; D012967:Sodium Dodecyl Sulfate; D004126:Dimethylformamide