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. 1992 Aug;174(16):5485–5487. doi: 10.1128/jb.174.16.5485-5487.1992

Membrane topology of the Escherichia coli ExbD protein.

K Kampfenkel 1, V Braun 1
PMCID: PMC206394  PMID: 1644779

Abstract

The ExbD protein is involved in the energy-coupled transport of ferric siderophores, vitamin B12, and B-group colicins across the outer membrane of Escherichia coli. In order to study ExbD membrane topology, ExbD-beta-lactamase fusion proteins were constructed. Cells expressing beta-lactamase fusions to residues 53, 57, 70, 76, 78, 80, 92, 121, and 134 of ExbD displayed high levels of ampicillin resistance, whereas fusions to residues 9 and 19 conferred no ampicillin resistance. It is concluded that the only hydrophobic segment of ExbD, encompassing residues 23 to 43, forms a transmembrane domain and that residues 1 to 22 are located in the cytoplasm and residues 44 to 141 are located in the periplasm.

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Selected References

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

  1. Anton M., Heller K. J. Functional analysis of a C-terminally altered TonB protein of Escherichia coli. Gene. 1991 Aug 30;105(1):23–29. doi: 10.1016/0378-1119(91)90509-a. [DOI] [PubMed] [Google Scholar]
  2. Braun V., Günter K., Hantke K. Transport of iron across the outer membrane. Biol Met. 1991;4(1):14–22. doi: 10.1007/BF01135552. [DOI] [PubMed] [Google Scholar]
  3. Braun V. The structurally related exbB and tolQ genes are interchangeable in conferring tonB-dependent colicin, bacteriophage, and albomycin sensitivity. J Bacteriol. 1989 Nov;171(11):6387–6390. doi: 10.1128/jb.171.11.6387-6390.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Broome-Smith J. K., Spratt B. G. A vector for the construction of translational fusions to TEM beta-lactamase and the analysis of protein export signals and membrane protein topology. Gene. 1986;49(3):341–349. doi: 10.1016/0378-1119(86)90370-7. [DOI] [PubMed] [Google Scholar]
  5. Broome-Smith J. K., Tadayyon M., Zhang Y. Beta-lactamase as a probe of membrane protein assembly and protein export. Mol Microbiol. 1990 Oct;4(10):1637–1644. doi: 10.1111/j.1365-2958.1990.tb00540.x. [DOI] [PubMed] [Google Scholar]
  6. Eick-Helmerich K., Braun V. Import of biopolymers into Escherichia coli: nucleotide sequences of the exbB and exbD genes are homologous to those of the tolQ and tolR genes, respectively. J Bacteriol. 1989 Sep;171(9):5117–5126. doi: 10.1128/jb.171.9.5117-5126.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Fischer E., Günter K., Braun V. Involvement of ExbB and TonB in transport across the outer membrane of Escherichia coli: phenotypic complementation of exb mutants by overexpressed tonB and physical stabilization of TonB by ExbB. J Bacteriol. 1989 Sep;171(9):5127–5134. doi: 10.1128/jb.171.9.5127-5134.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Grindley N. D., Joyce C. M. Genetic and DNA sequence analysis of the kanamycin resistance transposon Tn903. Proc Natl Acad Sci U S A. 1980 Dec;77(12):7176–7180. doi: 10.1073/pnas.77.12.7176. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Günter K., Braun V. In vivo evidence for FhuA outer membrane receptor interaction with the TonB inner membrane protein of Escherichia coli. FEBS Lett. 1990 Nov 12;274(1-2):85–88. doi: 10.1016/0014-5793(90)81335-l. [DOI] [PubMed] [Google Scholar]
  10. Hancock R. W., Braun V. Nature of the energy requirement for the irreversible adsorption of bacteriophages T1 and phi80 to Escherichia coli. J Bacteriol. 1976 Feb;125(2):409–415. doi: 10.1128/jb.125.2.409-415.1976. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Hannavy K., Barr G. C., Dorman C. J., Adamson J., Mazengera L. R., Gallagher M. P., Evans J. S., Levine B. A., Trayer I. P., Higgins C. F. TonB protein of Salmonella typhimurium. A model for signal transduction between membranes. J Mol Biol. 1990 Dec 20;216(4):897–910. doi: 10.1016/S0022-2836(99)80009-6. [DOI] [PubMed] [Google Scholar]
  12. Heller K. J., Kadner R. J., Günther K. Suppression of the btuB451 mutation by mutations in the tonB gene suggests a direct interaction between TonB and TonB-dependent receptor proteins in the outer membrane of Escherichia coli. Gene. 1988 Apr 15;64(1):147–153. doi: 10.1016/0378-1119(88)90488-x. [DOI] [PubMed] [Google Scholar]
  13. Hubacek J., Glover S. W. Complementation analysis of temperature-sensitive host specificity mutations in Escherichia coli. J Mol Biol. 1970 May 28;50(1):111–127. doi: 10.1016/0022-2836(70)90108-7. [DOI] [PubMed] [Google Scholar]
  14. Kadner R. J. Vitamin B12 transport in Escherichia coli: energy coupling between membranes. Mol Microbiol. 1990 Dec;4(12):2027–2033. doi: 10.1111/j.1365-2958.1990.tb00562.x. [DOI] [PubMed] [Google Scholar]
  15. Laemmli U. K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. doi: 10.1038/227680a0. [DOI] [PubMed] [Google Scholar]
  16. Mende J., Braun V. Import-defective colicin B derivatives mutated in the TonB box. Mol Microbiol. 1990 Sep;4(9):1523–1533. doi: 10.1111/j.1365-2958.1990.tb02063.x. [DOI] [PubMed] [Google Scholar]
  17. Mohana Rao J. K., Argos P. A conformational preference parameter to predict helices in integral membrane proteins. Biochim Biophys Acta. 1986 Jan 30;869(2):197–214. doi: 10.1016/0167-4838(86)90295-5. [DOI] [PubMed] [Google Scholar]
  18. Nau C. D., Konisky J. Evolutionary relationship between the TonB-dependent outer membrane transport proteins: nucleotide and amino acid sequences of the Escherichia coli colicin I receptor gene. J Bacteriol. 1989 Feb;171(2):1041–1047. doi: 10.1128/jb.171.2.1041-1047.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Nikaido H. Porins and specific channels of bacterial outer membranes. Mol Microbiol. 1992 Feb;6(4):435–442. doi: 10.1111/j.1365-2958.1992.tb01487.x. [DOI] [PubMed] [Google Scholar]
  20. Postle K. TonB and the gram-negative dilemma. Mol Microbiol. 1990 Dec;4(12):2019–2025. doi: 10.1111/j.1365-2958.1990.tb00561.x. [DOI] [PubMed] [Google Scholar]
  21. Reynolds P. R., Mottur G. P., Bradbeer C. Transport of vitamin B12 in Escherichia coli. Some observations on the roles of the gene products of BtuC and TonB. J Biol Chem. 1980 May 10;255(9):4313–4319. [PubMed] [Google Scholar]
  22. Roof S. K., Allard J. D., Bertrand K. P., Postle K. Analysis of Escherichia coli TonB membrane topology by use of PhoA fusions. J Bacteriol. 1991 Sep;173(17):5554–5557. doi: 10.1128/jb.173.17.5554-5557.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. 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]
  24. Schöffler H., Braun V. Transport across the outer membrane of Escherichia coli K12 via the FhuA receptor is regulated by the TonB protein of the cytoplasmic membrane. Mol Gen Genet. 1989 Jun;217(2-3):378–383. doi: 10.1007/BF02464907. [DOI] [PubMed] [Google Scholar]
  25. Skare J. T., Postle K. Evidence for a TonB-dependent energy transduction complex in Escherichia coli. Mol Microbiol. 1991 Dec;5(12):2883–2890. doi: 10.1111/j.1365-2958.1991.tb01848.x. [DOI] [PubMed] [Google Scholar]
  26. Studier F. W., Moffatt B. A. Use of bacteriophage T7 RNA polymerase to direct selective high-level expression of cloned genes. J Mol Biol. 1986 May 5;189(1):113–130. doi: 10.1016/0022-2836(86)90385-2. [DOI] [PubMed] [Google Scholar]
  27. Tabor S., Richardson C. C. A bacteriophage T7 RNA polymerase/promoter system for controlled exclusive expression of specific genes. Proc Natl Acad Sci U S A. 1985 Feb;82(4):1074–1078. doi: 10.1073/pnas.82.4.1074. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Van Hove B., Staudenmaier H., Braun V. Novel two-component transmembrane transcription control: regulation of iron dicitrate transport in Escherichia coli K-12. J Bacteriol. 1990 Dec;172(12):6749–6758. doi: 10.1128/jb.172.12.6749-6758.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Webster R. E. The tol gene products and the import of macromolecules into Escherichia coli. Mol Microbiol. 1991 May;5(5):1005–1011. doi: 10.1111/j.1365-2958.1991.tb01873.x. [DOI] [PubMed] [Google Scholar]

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