Biosynthesis and structure of lipopolysaccharide in an outer membrane-defective mutant of Escherichia coli J5.
Membrane-defective mutants of Escherichia coli J5 were isolated on the basis of supersensitivity to the antibiotic novobiocin. These mutants display an increased sensitivity to a wide range of antibiotics and to several dyes and detergents. In addition, several mutants leak the periplasmic enzymes, alkyline phosphatase and ribonuclease. This evidence indicates an outer membrane defect in these mutants. The inner and outer membranes of one mutant were separated and subjected to compositional analysis. A deficiency in galactose containing lipopolysaccharide in the outer membrane of the mutant was observed. Two possible causes of this deficiency were examined and discounted: defective galactose uptake into the cell, and defective translocation of lipopolysaccharide from the inner membrane. Extraction and chemical analysis of mutant and wild type lipopolysaccharides suggests that the mutant is defective in the enzyme which transfers glucose to the growing lipopolysaccharide core, UDPglucose transferase. Thus, the mutant's deficiency in galactose-containing lipopolysaccharide can be ascribed to the fact that addition of glucose to the lipopolysaccharide core is a prerequisite for galactose addition. The physiological implications of this alteration are discussed.
D001693:Biological Transport, Active; D002241:Carbohydrates / Q000032:analysis; D002458:Cell Fractionation; D002462:Cell Membrane / Q000187:drug effects / Q000378:metabolism*; D002499:Centrifugation, Density Gradient; D004926:Escherichia coli / Q000187:drug effects / Q000378:metabolism*; D005697:Galactosides / Q000378:metabolism; D006539:Heptoses / Q000378:metabolism; D007700:Kinetics; D008070:Lipopolysaccharides / Q000096:biosynthesis; D026901:Membrane Transport Proteins / Q000378:metabolism; D008968:Molecular Conformation; D009154:Mutation; D009675:Novobiocin / Q000494:pharmacology; D011135:Polysaccharides, Bacterial / Q000096:biosynthesis*
D002241:Carbohydrates; D005697:Galactosides; D006539:Heptoses; D008070:Lipopolysaccharides; D026901:Membrane Transport Proteins; D011135:Polysaccharides, Bacterial; D009675:Novobiocin