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. 1991 Sep;173(17):5554–5557. doi: 10.1128/jb.173.17.5554-5557.1991

Analysis of Escherichia coli TonB membrane topology by use of PhoA fusions.

S K Roof 1, J D Allard 1, K P Bertrand 1, K Postle 1
PMCID: PMC208271  PMID: 1885532

Abstract

Alkaline phosphatase (PhoA) fusions to TonB amino acids 32, 60, 125, 207, and 239 (the carboxy terminus) all showed high PhoA activity; a PhoA fusion to TonB amino acid 12 was inactive. The full-length TonB-PhoA fusion protein was associated with the cytoplasmic membrane and retained partial TonB function. These results support a model in which TonB is anchored in the cytoplasmic membrane by its hydrophobic amino terminus, with the remainder of the protein, including its hydrophobic carboxy terminus, extending into the periplasm.

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Selected References

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  1. Boyd D., Manoil C., Beckwith J. Determinants of membrane protein topology. Proc Natl Acad Sci U S A. 1987 Dec;84(23):8525–8529. doi: 10.1073/pnas.84.23.8525. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Calamia J., Manoil C. lac permease of Escherichia coli: topology and sequence elements promoting membrane insertion. Proc Natl Acad Sci U S A. 1990 Jul;87(13):4937–4941. doi: 10.1073/pnas.87.13.4937. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Devereux J., Haeberli P., Smithies O. A comprehensive set of sequence analysis programs for the VAX. Nucleic Acids Res. 1984 Jan 11;12(1 Pt 1):387–395. doi: 10.1093/nar/12.1part1.387. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Douthart R. J., Thomas J. J., Rosier S. D., Schmaltz J. E., West J. W. Cloning simulation in the cage environment. Nucleic Acids Res. 1986 Jan 10;14(1):285–297. doi: 10.1093/nar/14.1.285. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. 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]
  6. 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]
  7. 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]
  8. Kadner R. J., McElhaney G. Outer membrane-dependent transport systems in Escherichia coli: turnover of TonB function. J Bacteriol. 1978 Jun;134(3):1020–1029. doi: 10.1128/jb.134.3.1020-1029.1978. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Manoil C., Beckwith J. TnphoA: a transposon probe for protein export signals. Proc Natl Acad Sci U S A. 1985 Dec;82(23):8129–8133. doi: 10.1073/pnas.82.23.8129. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Michaelis S., Inouye H., Oliver D., Beckwith J. Mutations that alter the signal sequence of alkaline phosphatase in Escherichia coli. J Bacteriol. 1983 Apr;154(1):366–374. doi: 10.1128/jb.154.1.366-374.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Moyed H. S., Nguyen T. T., Bertrand K. P. Multicopy Tn10 tet plasmids confer sensitivity to induction of tet gene expression. J Bacteriol. 1983 Aug;155(2):549–556. doi: 10.1128/jb.155.2.549-556.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. 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]
  13. Plastow G. S., Holland I. B. Identification of an Escherichia coli inner membrane polypeptide specified by a lambda-tonB transducing. Biochem Biophys Res Commun. 1979 Oct 12;90(3):1007–1014. doi: 10.1016/0006-291x(79)91927-2. [DOI] [PubMed] [Google Scholar]
  14. Postle K., Good R. F. DNA sequence of the Escherichia coli tonB gene. Proc Natl Acad Sci U S A. 1983 Sep;80(17):5235–5239. doi: 10.1073/pnas.80.17.5235. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Postle K., Skare J. T. Escherichia coli TonB protein is exported from the cytoplasm without proteolytic cleavage of its amino terminus. J Biol Chem. 1988 Aug 5;263(22):11000–11007. [PubMed] [Google Scholar]
  16. 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]
  17. Pratt J. M., Jackson M. E., Holland I. B. The C terminus of penicillin-binding protein 5 is essential for localisation to the E. coli inner membrane. EMBO J. 1986 Sep;5(9):2399–2405. doi: 10.1002/j.1460-2075.1986.tb04510.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Skare J. T., Roof S. K., Postle K. A mutation in the amino terminus of a hybrid TrpC-TonB protein relieves overproduction lethality and results in cytoplasmic accumulation. J Bacteriol. 1989 Aug;171(8):4442–4447. doi: 10.1128/jb.171.8.4442-4447.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Towbin H., Staehelin T., Gordon J. Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications. Proc Natl Acad Sci U S A. 1979 Sep;76(9):4350–4354. doi: 10.1073/pnas.76.9.4350. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Wayne R., Neilands J. B. Evidence for common binding sites for ferrichrome compounds and bacteriophage phi 80 in the cell envelope of Escherichia coli. J Bacteriol. 1975 Feb;121(2):497–503. doi: 10.1128/jb.121.2.497-503.1975. [DOI] [PMC free article] [PubMed] [Google Scholar]

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