Skip to main content
Journal of Bacteriology logoLink to Journal of Bacteriology
. 1981 Apr;146(1):325–330. doi: 10.1128/jb.146.1.325-330.1981

Synthesis of a precursor to the B subunit of heat-labile enterotoxin in Escherichia coli.

E T Palva, T R Hirst, S J Hardy, J Holmgren, L Randall
PMCID: PMC217086  PMID: 6260742

Abstract

Escherichia coli K-12 minicells were employed to investigate the biosynthesis of plasmid-encoded, heat-labile enterotoxin of E. coli. Two polypeptide species related to the B subunit of the toxin were expressed in the minicells. One of these polypeptides (molecular weight, 11,500) was immunoprecipitated by antiserum to cholera toxin. Because the B subunits of heat-labile enterotoxin and cholera toxin have common antigenic sites, we concluded that this species was the mature B subunit. The larger polypeptide (molecular weight, 13,000) is likely to be a precursor of the B subunit because it could be chased into the mature form. This conversion was inhibited by compounds which dissipate proton motive force, suggesting that processing requires energy.

Full text

PDF
325

Images in this article

Selected References

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

  1. Cassel D., Pfeuffer T. Mechanism of cholera toxin action: covalent modification of the guanyl nucleotide-binding protein of the adenylate cyclase system. Proc Natl Acad Sci U S A. 1978 Jun;75(6):2669–2673. doi: 10.1073/pnas.75.6.2669. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Clements J. D., Finkelstein R. A. Isolation and characterization of homogeneous heat-labile enterotoxins with high specific activity from Escherichia coli cultures. Infect Immun. 1979 Jun;24(3):760–769. doi: 10.1128/iai.24.3.760-769.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Dallas W. S., Falkow S. Amino acid sequence homology between cholera toxin and Escherichia coli heat-labile toxin. Nature. 1980 Dec 4;288(5790):499–501. doi: 10.1038/288499a0. [DOI] [PubMed] [Google Scholar]
  4. Dallas W. S., Falkow S. The molecular nature of heat-labile enterotoxin (LT) of escherichia coli. Nature. 1979 Feb 1;277(5695):406–407. doi: 10.1038/277406a0. [DOI] [PubMed] [Google Scholar]
  5. Date T., Zwizinski C., Ludmerer S., Wickner W. Mechanisms of membrane assembly: effects of energy poisons on the conversion of soluble M13 coliphage procoat to membrane-bound coat protein. Proc Natl Acad Sci U S A. 1980 Feb;77(2):827–831. doi: 10.1073/pnas.77.2.827. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Dougan G., Saul M., Twigg A., Gill R., Sherratt D. Polypeptides expressed in Escherichia coli K-12 minicells by transposition elements Tn1 and Tn3. J Bacteriol. 1979 Apr;138(1):48–54. doi: 10.1128/jb.138.1.48-54.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Elder J. H., Pickett R. A., 2nd, Hampton J., Lerner R. A. Radioiodination of proteins in single polyacrylamide gel slices. Tryptic peptide analysis of all the major members of complex multicomponent systems using microgram quantities of total protein. J Biol Chem. 1977 Sep 25;252(18):6510–6515. [PubMed] [Google Scholar]
  8. Frazer A. C., Curtiss R., 3rd Production, properties and utility of bacterial minicells. Curr Top Microbiol Immunol. 1975;69:1–84. doi: 10.1007/978-3-642-50112-8_1. [DOI] [PubMed] [Google Scholar]
  9. Gyles C., So M., Falkow S. The enterotoxin plasmids of Escherichia coli. J Infect Dis. 1974 Jul;130(1):40–49. doi: 10.1093/infdis/130.1.40. [DOI] [PubMed] [Google Scholar]
  10. Harold F. M. Conservation and transformation of energy by bacterial membranes. Bacteriol Rev. 1972 Jun;36(2):172–230. doi: 10.1128/br.36.2.172-230.1972. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Hazelbauer G. L., Harayama S. Mutants in transmission of chemotactic signals from two independent receptors of E. coli. Cell. 1979 Mar;16(3):617–625. doi: 10.1016/0092-8674(79)90035-7. [DOI] [PubMed] [Google Scholar]
  12. Hindennach I., Henning U. The major proteins of the Excherichia coli outer cell envelope membrane. Preparative isolation of all major membrane proteins. Eur J Biochem. 1975 Nov 1;59(1):207–213. doi: 10.1111/j.1432-1033.1975.tb02443.x. [DOI] [PubMed] [Google Scholar]
  13. Inouye M., Halegoua S. Secretion and membrane localization of proteins in Escherichia coli. CRC Crit Rev Biochem. 1980;7(4):339–371. doi: 10.3109/10409238009105465. [DOI] [PubMed] [Google Scholar]
  14. Kunkel S. L., Robertson D. C. Purification and chemical characterization of the heat-labile enterotoxin produced by enterotoxigenic Escherichia coli. Infect Immun. 1979 Aug;25(2):586–596. doi: 10.1128/iai.25.2.586-596.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Lai C. Y., Cancedda F., Chang D. Primary structure of cholera toxin subunit A1: isolation, partial sequences and alignment of the BrCN fragments. FEBS Lett. 1979 Apr 1;100(1):85–89. doi: 10.1016/0014-5793(79)81136-9. [DOI] [PubMed] [Google Scholar]
  16. Lai C. Y. Determination of the primary structure of cholera toxin B subunit. J Biol Chem. 1977 Oct 25;252(20):7249–7256. [PubMed] [Google Scholar]
  17. Larsen S. H., Adler J., Gargus J. J., Hogg R. W. Chemomechanical coupling without ATP: the source of energy for motility and chemotaxis in bacteria. Proc Natl Acad Sci U S A. 1974 Apr;71(4):1239–1243. doi: 10.1073/pnas.71.4.1239. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Randall L. L., Josefsson L. G., Hardy S. J. Processing in vitro of precursor periplasmic proteins from Escherichia coli. Eur J Biochem. 1978 Dec;92(2):411–415. doi: 10.1111/j.1432-1033.1978.tb12761.x. [DOI] [PubMed] [Google Scholar]
  19. So M., Dallas W. S., Falkow S. Characterization of an Escherichia coli plasmid encoding for synthesis of heat-labile toxin: molecular cloning of the toxin determinant. Infect Immun. 1978 Aug;21(2):405–411. doi: 10.1128/iai.21.2.405-411.1978. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from Journal of Bacteriology are provided here courtesy of American Society for Microbiology (ASM)

RESOURCES