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. 1983 Sep;155(3):1382–1392. doi: 10.1128/jb.155.3.1382-1392.1983

Monoclonal antibodies reveal lamB antigenic determinants on both faces of the Escherichia coli outer membrane.

S Schenkman, E Couture, M Schwartz
PMCID: PMC217838  PMID: 6193098

Abstract

LamB protein is involved in the transport of maltose across the outer membrane and constitutes the receptor for phage lambda. In this study we characterized six previously described anti-LamB monoclonal antibodies (mAbs). Four of these, the E-mAbs, recognized determinants that were exposed at the cell surface, whereas the other two, the I-mAbs, recognized determinants which were not exposed. Competition experiments demonstrated that the domains recognized by these two classes of mAbs were completely distinct. In addition, the E-mAbs prevented LamB from neutralizing phage lambda in vitro and protected LamB against proteolytic degradation, whereas the I-mAbs had no such effects. The E-mAbs have been shown previously to constitute two classes: some E-mAbs inhibit maltose transport in vivo, and others do not. Immunoelectron microscopy demonstrated that the I-mAbs also define at least two types of determinants. One of these, which is accessible in membrane fragments from a mutant (lpp) devoid of lipoprotein but not in membrane fragments from an lpp+ strain, probably corresponds to a region of LamB that is involved in the interactions with peptidoglycan. The other determinant, which is fully accessible in LamB-peptidoglycan complexes and in LamB-containing phospholipid vesicles but only slightly accessible in membrane fragments from an lpp mutant, is probably located very close to the inner surface of the outer membrane. LamB also contains at least one additional determinant, which (i) is exposed at the inner surface of the membrane, (ii) is accessible to antibodies in membrane fragments from an lpp+ strain, and (iii) may be involved in the interaction of LamB with the periplasmic maltose-binding protein.

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Bavoil P., Nikaido H. Physical interaction between the phage lambda receptor protein and the carrier-immobilized maltose-binding protein of Escherichia coli. J Biol Chem. 1981 Nov 25;256(22):11385–11388. [PubMed] [Google Scholar]
  2. Braun V., Krieger-Brauer H. J. Interrelationship of the phage lambda receptor protein and maltose transport in mutants of Escherichia coli K12. Biochim Biophys Acta. 1977 Aug 15;469(1):89–98. doi: 10.1016/0005-2736(77)90328-5. [DOI] [PubMed] [Google Scholar]
  3. 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. Eur J Biochem. 1969 Oct;10(3):426–438. doi: 10.1111/j.1432-1033.1969.tb00707.x. [DOI] [PubMed] [Google Scholar]
  4. Casadaban M. J. Transposition and fusion of the lac genes to selected promoters in Escherichia coli using bacteriophage lambda and Mu. J Mol Biol. 1976 Jul 5;104(3):541–555. doi: 10.1016/0022-2836(76)90119-4. [DOI] [PubMed] [Google Scholar]
  5. Clément J. M., Hofnung M. Gene sequence of the lambda receptor, an outer membrane protein of E. coli K12. Cell. 1981 Dec;27(3 Pt 2):507–514. doi: 10.1016/0092-8674(81)90392-5. [DOI] [PubMed] [Google Scholar]
  6. Ferenci T., Schwentorat M., Ullrich S., Vilmart J. Lambda receptor in the outer membrane of Escherichia coli as a binding protein for maltodextrins and starch polysaccharides. J Bacteriol. 1980 May;142(2):521–526. doi: 10.1128/jb.142.2.521-526.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Gabay J., Benson S., Schwartz M. Genetic mapping of antigenic determinants on a membrane protein. J Biol Chem. 1983 Feb 25;258(4):2410–2414. [PubMed] [Google Scholar]
  8. Gabay J., Schwartz M. Monoclonal antibody as a probe for structure and function of an Escherichia coli outer membrane protein. J Biol Chem. 1982 Jun 25;257(12):6627–6630. [PubMed] [Google Scholar]
  9. Gabay J., Yasunaka K. Interaction of the lamB protein with the peptidoglycan layer in Escherichia coli K12. Eur J Biochem. 1980 Feb;104(1):13–18. doi: 10.1111/j.1432-1033.1980.tb04393.x. [DOI] [PubMed] [Google Scholar]
  10. Geoghegan W. D., Ackerman G. A. Adsorption of horseradish peroxidase, ovomucoid and anti-immunoglobulin to colloidal gold for the indirect detection of concanavalin A, wheat germ agglutinin and goat anti-human immunoglobulin G on cell surfaces at the electron microscopic level: a new method, theory and application. J Histochem Cytochem. 1977 Nov;25(11):1187–1200. doi: 10.1177/25.11.21217. [DOI] [PubMed] [Google Scholar]
  11. Heuzenroeder M. W., Reeves P. Periplasmic maltose-binding protein confers specificity on the outer membrane maltose pore of Escherichia coli. J Bacteriol. 1980 Feb;141(2):431–435. doi: 10.1128/jb.141.2.431-435.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Hofnung M., Jezierska A., Braun-Breton C. lamB mutations in E. coli K12: growth of lambda host range mutants and effect of nonsense suppressors. Mol Gen Genet. 1976 May 7;145(2):207–213. doi: 10.1007/BF00269595. [DOI] [PubMed] [Google Scholar]
  13. Ishii J. N., Okajima Y., Nakae T. Characterization of lamB protein from the outer membrane of Escherichia coli that forms diffusion pores selective for maltose-maltodextrins. FEBS Lett. 1981 Nov 16;134(2):217–220. doi: 10.1016/0014-5793(81)80605-9. [DOI] [PubMed] [Google Scholar]
  14. Ito K., Date T., Wickner W. Synthesis, assembly into the cytoplasmic membrane, and proteolytic processing of the precursor of coliphage M13 coat protein. J Biol Chem. 1980 Mar 10;255(5):2123–2130. [PubMed] [Google Scholar]
  15. LOWRY O. H., ROSEBROUGH N. J., FARR A. L., RANDALL R. J. Protein measurement with the Folin phenol reagent. J Biol Chem. 1951 Nov;193(1):265–275. [PubMed] [Google Scholar]
  16. Luckey M., Nikaido H. Bacteriophage lambda receptor protein in Escherichia coli K-12: lowered affinity of some mutant proteins for maltose-binding protein in vitro. J Bacteriol. 1983 Feb;153(2):1056–1059. doi: 10.1128/jb.153.2.1056-1059.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Luckey M., Nikaido H. Specificity of diffusion channels produced by lambda phage receptor protein of Escherichia coli. Proc Natl Acad Sci U S A. 1980 Jan;77(1):167–171. doi: 10.1073/pnas.77.1.167. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Marchal C., Hofnung M. Negative dominance in gene lamB: random assembly of secreted subunits issued from different polysomes. EMBO J. 1983;2(1):81–86. doi: 10.1002/j.1460-2075.1983.tb01385.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Neuhaus J. M. The receptor protein of phage lambda: purification, characterization and preliminary electrical studies in planar lipid bilayers. Ann Microbiol (Paris) 1982 Jan;133A(1):27–32. [PubMed] [Google Scholar]
  20. Osborn M. J., Gander J. E., Parisi E., Carson J. Mechanism of assembly of the outer membrane of Salmonella typhimurium. Isolation and characterization of cytoplasmic and outer membrane. J Biol Chem. 1972 Jun 25;247(12):3962–3972. [PubMed] [Google Scholar]
  21. Osborn M. J., Wu H. C. Proteins of the outer membrane of gram-negative bacteria. Annu Rev Microbiol. 1980;34:369–422. doi: 10.1146/annurev.mi.34.100180.002101. [DOI] [PubMed] [Google Scholar]
  22. Palva E. T. Protein interactions in the outer membrane of Escherichia coli. Eur J Biochem. 1979 Feb 1;93(3):495–503. doi: 10.1111/j.1432-1033.1979.tb12848.x. [DOI] [PubMed] [Google Scholar]
  23. Randall-Hazelbauer L., Schwartz M. Isolation of the bacteriophage lambda receptor from Escherichia coli. J Bacteriol. 1973 Dec;116(3):1436–1446. doi: 10.1128/jb.116.3.1436-1446.1973. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Roa M. Interaction of bacteriophage K10 with its receptor, the lamB protein of Escherichia coli. J Bacteriol. 1979 Nov;140(2):680–686. doi: 10.1128/jb.140.2.680-686.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Schwartz M., Le Minor L. Occurrence of the bacteriophage lambda receptor in some enterobacteriaceae. J Virol. 1975 Apr;15(4):679–685. doi: 10.1128/jvi.15.4.679-685.1975. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Shuman H. A. The maltose-maltodextrin transport system of Escherichia coli. Ann Microbiol (Paris) 1982 Jan;133A(1):153–159. [PubMed] [Google Scholar]
  27. Suzuki H., Nishimura Y., Yasuda S., Nishimura A., Yamada M., Hirota Y. Murein-lipoprotein of Escherichia coli: a protein involved in the stabilization of bacterial cell envelope. Mol Gen Genet. 1978 Nov 16;167(1):1–9. doi: 10.1007/BF00270315. [DOI] [PubMed] [Google Scholar]
  28. Szmelcman S., Hofnung M. Maltose transport in Escherichia coli K-12: involvement of the bacteriophage lambda receptor. J Bacteriol. 1975 Oct;124(1):112–118. doi: 10.1128/jb.124.1.112-118.1975. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Wandersman C., Schwartz M., Ferenci T. Escherichia coli mutants impaired in maltodextrin transport. J Bacteriol. 1979 Oct;140(1):1–13. doi: 10.1128/jb.140.1.1-13.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Yamada H., Nogami T., Mizushima S. Arrangement of bacteriophage lambda receptor protein (LamB) in the cell surface of Escherichia coli: a reconstitution study. J Bacteriol. 1981 Aug;147(2):660–669. doi: 10.1128/jb.147.2.660-669.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]

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