Cellular location of heat-labile enterotoxin in Escherichia coli

Hirst, Timothy R.; Randall, Linda L.; Hardy, Simon

doi:10.1128/jb.157.2.637-642.1984

Cited by 91 publications

(21 citation statements)

References 37 publications

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“…Heat-labile enterotoxin (HLET) of E. coli, and the closely related cholera toxin (CT) of Vibrio cholerae, are synthesised as two subunits, each of which initially carries a typical NH2-terminal signal peptide [270,271,297,300,[342][343][344][345]. Both subunits are apparently required for their release from producing cells [346].…”

Section: Heat-labile Enterotoxin and Cholera Toxinmentioning

confidence: 99%

Export and secretion of proteins by bacteria

Pugsley

Schwartz

1985

FEMS Microbiology Letters

237

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A wide variety of proteins are exported or secreted by a range of morphologically distinct bacteria. The processes of protein export are most extensively characterised in Escherichia coli, where recent advances have been made in the identification of genes involved in forming the export machinery. Both Gram‐positive and Gram‐negative bacteria secrete proteins into the medium. Gram‐negative bacteria have adopted a variety of approaches in order to overcome the additional permeability of the outer membrane.

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Section: Heat-labile Enterotoxin and Cholera Toxinmentioning

confidence: 99%

Export and secretion of proteins by bacteria

Pugsley

Schwartz

1985

FEMS Microbiology Letters

237

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“…Periplasmic fractions of E. coli were prepared by the method of Hirst et al (1984). Proteins were subsequently visualized by staining with Coomassie Brilliant Blue or electrobiotted onto nitrocellulose and probed with polyclonal rabbit anti-LTB antisera.…”

Section: Preparation and Analysis Of Periplasmic Extractsmentioning

confidence: 99%

Intranasal immunization using the B subunit of the Escherichia coli heat‐labile toxin fused to an epitope of the Bordetella pertussis P.69 antigen

Lipscombe

Roberts

et al. 1991

Molecular Microbiology

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The plasmid pBRD026, which directs expression of the B subunit of the Escherichia coli heat-labile toxin (LTB), was modified so that DNA encoding epitopes could be inserted at the 3' end of the gene. An oligonucleotide linker containing restriction sites for BglII and SpeI was inserted at the SpeI site at the 3' end of the LTB gene to form plasmid pFV1. This linker also encodes the amino acid sequence Gly-Pro-Gly-Pro which we propose acts as a 'hinge' between the LTB and the foreign epitope. Oligonucleotides specifying an epitope from the Bordetella pertussis P.69 outer membrane protein were cloned into pFV1 to form pFV169. The resultant fusion protein (LTB69) was partially purified from the periplasm of E. coli strains in a soluble pentameric form which could bind GM1 gangliosides. Mice immunized intranasally with purified LTB69 produced antibodies against both LTB and the P.69 protein. In addition, ELISPOT assays demonstrated the presence of LTB-specific and P.69-specific antibody-secreting cells in the lungs of immunized mice.

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“…Polypeptide 115 fractionated approximately equaify between ttie envelope and soluble fraction whilst poiypeptide 116 and authentic PBP5 fractionated mainly with the envelope fraction. To confirm that membrane association rather than formation of protein aggregates was responsible for the presence of the internally deleted PBP5 molecules in the envelope fraction, the membranes were floated up through sucrose gradients (Hirst et al, 1984), As shown in Fig. 5.…”

Section: Localization Of Internally Deleted Pbp5 Moleculesmentioning

confidence: 99%

An 18 amino acid amphiphilic helix forms the membrane‐anchoring domain of the Escherichia coli penicillin‐binding protein 5

Jackson

Pratt

1987

Molecular Microbiology

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Small (10 residue) C-terminal deletions of PBP5 cause release of this inner membrane protein into the periplasm, indicating disruption of the membrane binding domain. To define the extent of the membrane anchoring domain, oligonucleotide-directed mutagenesis was used to introduce both single amino acid changes and novel restriction sites into the DNA, allowing subsequent construction of precise internal deletions. The 10 C-terminal amino acid residues possess very weak membrane anchoring potential. By extending the sequence to 18 residues membrane binding equivalent to that of authentic PBP5 was achieved. A proline substitution in this region, breaking a potential alpha-helix, also disrupts the membrane binding domain. These results are discussed with respect to the amphiphilicity of the C-terminal sequence when arranged in an alpha-helix.

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Cellular location of heat-labile enterotoxin in Escherichia coli

Cited by 91 publications

References 37 publications

Export and secretion of proteins by bacteria

Export and secretion of proteins by bacteria

Intranasal immunization using the B subunit of the Escherichia coli heat‐labile toxin fused to an epitope of the Bordetella pertussis P.69 antigen

An 18 amino acid amphiphilic helix forms the membrane‐anchoring domain of the Escherichia coli penicillin‐binding protein 5

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