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. 1986 Jun;5(6):1383–1388. doi: 10.1002/j.1460-2075.1986.tb04371.x

Mutations affecting antigenic determinants of an outer membrane protein of Escherichia coli.

C Desaymard, M Débarbouillé, M Jolit, M Schwartz
PMCID: PMC1166952  PMID: 2426104

Abstract

The Escherichia coli LamB protein is located in the outer membrane. It is both a component of the maltose and maltodextrin transport system, and the receptor for phages lambda and K10. It is a trimer composed of three identical polypeptide chains, each containing 421 residues. Six independent mutants have been isolated, in which the LamB protein is altered in its interaction with one or more monoclonal antibodies specific for regions of the protein that are exposed at the cell surface. Some of the mutations also altered the binding site for phage lambda. All of the mutations were clustered in the same region of the lamB gene, corresponding to residues 333-394 in the polypeptide. This and previous results strongly suggest that a rather large segment of the LamB polypeptide, extending from residue 315 to 401, is exposed at the outer face of the outer membrane. This segment would bear the epitopes for the four available anti-LamB monoclonal antibodies that react with the cell surface, and part of the binding site for phage lambda.

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Selected References

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

  1. Bloch M. A., Desaymard C. Antigenic polymorphism of the LamB protein among members of the family Enterobacteriaceae. J Bacteriol. 1985 Jul;163(1):106–110. doi: 10.1128/jb.163.1.106-110.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. 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]
  3. Charbit A., Clement J. M., Hofnung M. Further sequence analysis of the phage lambda receptor site. Possible implications for the organization of the lamB protein in Escherichia coli K12. J Mol Biol. 1984 May 25;175(3):395–401. doi: 10.1016/0022-2836(84)90355-3. [DOI] [PubMed] [Google Scholar]
  4. Chattoraj D. K., Cordes K., Berman M. L., Das A. Mutagenesis and mutation transfer induced by ultraviolet light in plasmid-cloned DNA. Gene. 1984 Feb;27(2):213–222. doi: 10.1016/0378-1119(84)90142-2. [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. Clément J. M., Lepouce E., Marchal C., Hofnung M. Genetic study of a membrane protein: DNA sequence alterations due to 17 lamB point mutations affecting adsorption of phage lambda. EMBO J. 1983;2(1):77–80. doi: 10.1002/j.1460-2075.1983.tb01384.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Deisenhofer J., Epp O., Miki K., Huber R., Michel H. X-ray structure analysis of a membrane protein complex. Electron density map at 3 A resolution and a model of the chromophores of the photosynthetic reaction center from Rhodopseudomonas viridis. J Mol Biol. 1984 Dec 5;180(2):385–398. doi: 10.1016/s0022-2836(84)80011-x. [DOI] [PubMed] [Google Scholar]
  8. 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]
  9. 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]
  10. Henderson R., Unwin P. N. Three-dimensional model of purple membrane obtained by electron microscopy. Nature. 1975 Sep 4;257(5521):28–32. doi: 10.1038/257028a0. [DOI] [PubMed] [Google Scholar]
  11. 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]
  12. 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]
  13. 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]
  14. Palva E. T., Westermann P. Arrangement of the maltose-inducible major outer membrane proteins, the bacteriophage lambda receptor in Escherichia coli and the 44 K protein in Salmonella typhimurium. FEBS Lett. 1979 Mar 1;99(1):77–80. doi: 10.1016/0014-5793(79)80253-7. [DOI] [PubMed] [Google Scholar]
  15. Raibaud O., Roa M., Braun-Breton C., Schwartz M. Structure of the malB region in Escherichia coli K12. I. Genetic map of the malK-lamB operon. Mol Gen Genet. 1979 Jul 24;174(3):241–248. doi: 10.1007/BF00267796. [DOI] [PubMed] [Google Scholar]
  16. Roa M., Clément J. M. Location of a phage binding region on an outer membrane protein. FEBS Lett. 1980 Nov 17;121(1):127–129. doi: 10.1016/0014-5793(80)81280-4. [DOI] [PubMed] [Google Scholar]
  17. Sadler J. R., Tecklenburg M., Betz J. L., Goeddel D. V., Yansura D. G., Caruthers M. H. Cloning of chemically synthesized lactose operators. Gene. 1977 Jul;1(5-6):305–321. doi: 10.1016/0378-1119(77)90036-1. [DOI] [PubMed] [Google Scholar]
  18. Sanger F., Nicklen S., Coulson A. R. DNA sequencing with chain-terminating inhibitors. Proc Natl Acad Sci U S A. 1977 Dec;74(12):5463–5467. doi: 10.1073/pnas.74.12.5463. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Schenkman S., Couture E., Schwartz M. Monoclonal antibodies reveal lamB antigenic determinants on both faces of the Escherichia coli outer membrane. J Bacteriol. 1983 Sep;155(3):1382–1392. doi: 10.1128/jb.155.3.1382-1392.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Schenkman S., Tsugita A., Schwartz M., Rosenbusch J. P. Topology of phage lambda receptor protein. Mapping targets of proteolytic cleavage in relation to binding sites for phage or monoclonal antibodies. J Biol Chem. 1984 Jun 25;259(12):7570–7576. [PubMed] [Google Scholar]
  21. 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]
  22. Wandersman C., Schwartz M. Mutations that alter the transport function of the LamB protein in Escherichia coli. J Bacteriol. 1982 Jul;151(1):15–21. doi: 10.1128/jb.151.1.15-21.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]

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