Summary
Two independent mutants of Escherichia coli lacking murein-lipoprotein have been found. One mutant whose mutation was named lpo was subjected to detailed analyses. The absence of both found and unbound lipoproteins was shown by electrophoretic analysis of 14C-arginine labelled membrane proteins of the mutant. Nor was serologically cross-reacting material detected in the mutant by the Ouchterlony-method. Sequestering magnesium from mutant cell suspensions by ethylenediaminetetraacetic acid caused cell lysis, which was prevented in the presence of 0.5 M sucrose. Incubation in culture media at a very low level of magnesium resulted in the formation of blebs in the mutant. Examination of mutant cells by electron microscopy showed that the outer membrane of the mutant was uneven with small irregular protuberances, some of which pinched off forming vesicles of various sizes. Phosphotungstate used for negative-staining penetrated into the periplasmic space of the mutant cells. The mutants leaked a considerable fraction of their periplasmic enzymes. These physiological and morphological alterations in the lipoproteinless mutant suggest that murein-lipoprotein helps to maintain the outer envelope structure by connecting the outer membrane with murein so that the outer membrane may fulfil its physiological functions as a barrier to the environment.
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References
Anraku, Y.: Cyclic phosphodiesterase of Escherichia coli. In: Procedures in nucleic acid research (Cantoni, G.L. and Davies, D.R. eds), Vol. 1. p. 130. New York: Harper & Row 1966
Bachmann, B.J., Low, K.B., Taylor, A.L.: Recalibrated linkage map of Escherichia coli K-12. Bact. Rev. 40, 116–167 (1976)
Braun, V.: Covalent lipoprotein from the outer membrane of Escherichia coli. Biochim. biophys. Acta (Amst.) 415, 335–377 (1975)
Braun, V., Bosch, V.: Repetitive sequences in the murein-lipoprotein of the cell wall of Escherichia coli. Proc. nat. Acad. Sci. (Wash.) 69, 970–974 (1972)
Braun, V., Hantke, K.: Biochemistry of bacterial cell envelopes. Ann. Rev. Biochem. 43, 89–121 (1974)
Braun, V., Rehn, K.: Chemical characterization, spatial distribution and function of a lipoprotein (murein-lipoprotein) of the E. coli cell wall. The specific effect of trypsin on the membrane structure. Europ. J. Biochem. 10, 426–438 (1969)
Braun, V., Rehn, K., Walff, H.: Supramolecular structure of the rigid layer of the cell wall of Salmonella, Serratia, Proteus and Pseudomonas fluorescens Number of lipoprotein molecules in membrane layer. Biochemistry (Wash.) 22, 5041–5049 (1970)
Davis, B.D., Mingioli, E.S.: Mutants of Escherichia coli requiring methionine or vitamin B12. J. Bact. 60, 17–28 (1950)
Garen, A., Levinthal, C.: A fine-structure genetic and chemical study of the enzyme alkaline phosphatase of E. coli. I. Purification and characterization of alkaline phosphatase. Biochim. biophys. Acta (Amst.) 38, 478–483 (1960)
Halegoua, S., Hirashima, A., Inouye, M.: Existence of a free form of a specific membrane lipoprotein in gram negative bacteria. J. Bact. 120, 1204–1208 (1974)
Hantke, K., Braun, V.: Covalent binding of lipid to protein. Diglyceride and amide-linked fatty acid at the N-terminal end of the murein-lipoprotein of the Escherichia coli outer membrane. Europ. J. Biochem. 34, 284–296 (1973)
Hasegawa, Y., Yamada, H., Mizushima, S.: Interactions of outer membrane proteins-O-8 and O-9 with peptidoglycan sacculus of Escherichia coli K-12. J. Biochem. (Tokyo) 80, 1401–1409 (1976)
Henning, U., Rehn, K., Braun, V., Höhn, B., Schwarz, U.: Cell envelope and shape of Escherichia coli K12. Properties of a temperature-sensitive rod mutant. Europ. J. Biochem. 26, 570–586 (1972)
Hirota, Y., Suzuki, H., Nishimura, Y., Yasuda, S.: On the process of cellular division in Escherichia coli: A mutant of E. coli lacking a murein-lipoprotein. Proc. nat. Acad. Sci. (Wash.) 74, 1417–1420 (1977)
Inouye, M.: A three-dimensional molecular assembly model of a lipoprotein from the Escherichia coli outer membrane. Proc. nat. Acad. Sci. (Wash.) 71, 2396–2400 (1974)
Inouye, M., Guthrie, J.P.: A mutation which changes a membrane protein of E. coli. Proc. nat. Acad. Sci. (Wash.) 64, 957–961 (1969)
Inouye, M., Shaw, J., Shen, C.: The assembly of a structural lipoprotein in the envelope of Escherichia coli. J. biol. Chem. 246, 8154–8159 (1972)
Inouye, S., Lee, N., Inouye, M., Wu, H.C., Suzuki, H., Nishimura, Y., Iketani, H., Hirota, Y.: Amino acid replacement in a mutant lipoprotein of the outer membrane of Escherichia coli. J. Bact. 132, 308–313 (1977)
Kellenberger, E., Ryter, A., Sechaud, J.: Electron microscope study of DNA containing plasmids. II. Vegetative and mature phage DNA as compared in different physiological states. J. biophys. biochem. Cytol. 4, 671–678 (1958)
Lennox, E.S.: Transduction of linked genetic characters of the host by bacteriophage P1. Virology 1, 190–206 (1955)
LePage, G.A.: Methods for the analysis of phosphorylated intermediates. In: Manometric techniques (Umbreit, W.W., Burris, R.H. and Stanffer, J.F., eds.), p. 268. Minneapolis. Burgess Publ. Co. 1957
Matsuhashi, M., Takagaki, Y., Maruyama, I.N., Tamaki, S., Nishimura, Y., Suzuki, H., Ogino, U., Hirota, Y.: Mutants of Escherichia coli lacking in highly penicillin-sensitive D-alanine carboxypeptidase activity. Proc. nat. Acad. Sci. (Wash.) 74, 2976–2979 (1977)
Nikaido, H.: Biosynthesis and assembly of lipopolysacchride and the outer membrane layer of gram-negative cell wall. In: Bacterial membranes and walls (Laskin, A.I., ed.), Vol. 1, p. 131. New York, Marcel Dekker Inc. 1973
Platt, T., Müller-Hill, B., Miller, J.H.: Assay of β-galactosidase. In: Experiments in molecular genetics, p. 352. Cold Spring Harbor, New York: Cold Spring Harbor Laboratory 1972
Spahr, P.F.: The isolation, assay, and properties of ribonuclease I from Escherichia coli. In: Procedures in nucleic acid research (Cantoni, G.L. and Davies, D.R. eds.), Vol. 1, p. 64. New York: Harper & Row 1966
Suzuki, H., Iino, T.: In vitro synthesis of phase-specific flagellin of Salmonella. J. molec. Biol. 81, 57–70 (1973)
Suzuki, H., Nishimura, Y., Iketani, H., Campisi, J., Hirashima, A., Inouye, M., Hirota, Y.: Novel mutation that causes a structural changes in a lipoprotein in the outer membrane of Escherichia coli. J. Bact. 127, 1494–1501 (1976)
Torti, S.V., Park, J.I.: Lipoprotein of gram-negative bacteria is essential for growth and division. Nature (Lond.) 263, 323–326 (1976)
Voll, M.J., Leive, L.: Release of lipopolysaccharide in Escherichia coli resistant to the permeability increase induced by ethylenediaminetetraacetate. J. biol. Chem. 245, 1108–1114 (1970)
Weidel, W., Pelzer, H.: Bagshaped macromolecules: a new outlook on bacterial cell walls. Advanc. Enzymol. 26, 193–232 (1964)
Weigand, R.A., Vinch, K.D., Rothfield, L.I.: Morphogenesis of the bacterial division septum: a new class of septation-defective mutants. Proc. nat. Acad. Sci. (Wash.) 73, 1882–1886 (1976)
Wu, H.C., Lin, J.J.: Escherichia coli mutant altered in murein lipoprotein. J. Bact. 126, 147–156 (1976)
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Suzuki, H., Nishimura, Y., Yasuda, S. et al. Murein-lipoprotein of Escherichia coli: A protein involved in the stabilization of bacterial cell envelope. Molec. Gen. Genet. 167, 1–9 (1978). https://doi.org/10.1007/BF00270315
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DOI: https://doi.org/10.1007/BF00270315