Skip to main content
PMC home page
Infection and Immunity logoLink to Infection and Immunity
. 1981 Sep;33(3):877–883. doi: 10.1128/iai.33.3.877-883.1981

Purification, characterization, and partial covalent structure of Escherichia coli adhesive antigen K99.

F K de Graaf, P Klemm, W Gaastra
PMCID: PMC350793  PMID: 7026448

Abstract

The adhesive antigen K99 of enterotoxigenic Escherichia coli strains of calf origin was isolated and purified. The K99 fimbriae were removed from the cells by heat treatment, concentrated by precipitation with ammonium sulfate, and purified by gel filtration on Sepharose CL-4B and treatment with deoxycholate. The purified K99 antigen was composed of protein subunits with a molecular weight of 18,500 and had an isoelectric point of 9.5. The N-terminal amino acid sequence, as well as the composition of the C-terminal part of the K99 protein subunits, was determined.

Full text

PDF
877

Images in this article

879Image
on p.879
880Image
on p.880
882Image
on p.882

Selected References

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

  1. Evans D. G., Evans D. J., Jr, Clegg S., Pauley J. A. Purification and characterization of the CFA/I antigen of enterotoxigenic Escherichia coli. Infect Immun. 1979 Aug;25(2):738–748. doi: 10.1128/iai.25.2.738-748.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Evans D. G., Evans D. J., Jr New surface-associated heat-labile colonization factor antigen (CFA/II) produced by enterotoxigenic Escherichia coli of serogroups O6 and O8. Infect Immun. 1978 Aug;21(2):638–647. doi: 10.1128/iai.21.2.638-647.1978. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Evans D. G., Evans D. J., Jr, Tjoa W. S., DuPont H. L. Detection and characterization of colonization factor of enterotoxigenic Escherichia coli isolated from adults with diarrhea. Infect Immun. 1978 Feb;19(2):727–736. doi: 10.1128/iai.19.2.727-736.1978. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Guinée P. A., Jansen W. H., Agterberg C. M. Detection of the K99 antigen by means of agglutination and immunoelectrophoresis in Escherichia coli isolates from calves and its correlation with entertoxigenicity. Infect Immun. 1976 May;13(5):1369–1377. doi: 10.1128/iai.13.5.1369-1377.1976. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Hermodson M. A., Chen K. C., Buchanan T. M. Neisseria pili proteins: amino-terminal amino acid sequences and identification of an unusual amino acid. Biochemistry. 1978 Feb 7;17(3):442–445. doi: 10.1021/bi00596a010. [DOI] [PubMed] [Google Scholar]
  6. Isaacson R. E. K99 surface antigen of Escherichia coli: antigenic characterization. Infect Immun. 1978 Nov;22(2):555–559. doi: 10.1128/iai.22.2.555-559.1978. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Isaacson R. E. K99 surface antigen of Escherichia coli: purification and partial characterization. Infect Immun. 1977 Jan;15(1):272–279. doi: 10.1128/iai.15.1.272-279.1977. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Klemm P. Fimbrial colonization factor CFA/1 protein from human enteropathogenic Escherichia coli strains. Purification, characterization and N-terminal sequence. FEBS Lett. 1979 Dec 1;108(1):107–110. doi: 10.1016/0014-5793(79)81188-6. [DOI] [PubMed] [Google Scholar]
  9. Klemm P., Poulsen F., Harboe M. K., Foltmann B. N-terminal amino acid sequences of pepsinogens from dogfish and seal and of bovine pepsinogen B. Acta Chem Scand B. 1976;30(10):979–984. doi: 10.3891/acta.chem.scand.30b-0979. [DOI] [PubMed] [Google Scholar]
  10. Korhonen T. K., Nurmiaho E. L., Ranta H., Edén C. S. New Method for isolation of immunologically pure pili from Escherichia coli. Infect Immun. 1980 Feb;27(2):569–575. doi: 10.1128/iai.27.2.569-575.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Lugtenberg B., Meijers J., Peters R., van der Hoek P., van Alphen L. Electrophoretic resolution of the "major outer membrane protein" of Escherichia coli K12 into four bands. FEBS Lett. 1975 Oct 15;58(1):254–258. doi: 10.1016/0014-5793(75)80272-9. [DOI] [PubMed] [Google Scholar]
  12. Mooi F. R., de Graaf F. K., van Embden J. D. Cloning, mapping and expression of the genetic determinant that encodes for the K88ab antigen. Nucleic Acids Res. 1979 Mar;6(3):849–865. doi: 10.1093/nar/6.3.849. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Morris J. A., Stevens A. E., Sojka W. J. Anionic and cationic components of the K99 surface antigen from Escherichia coli B41. J Gen Microbiol. 1978 Jul;107(1):173–175. doi: 10.1099/00221287-107-1-173. [DOI] [PubMed] [Google Scholar]
  14. Morris J. A., Stevens A. E., Sojka W. J. Isoelectric point of cell-free K99 antigen exhibiting hemagglutinating properties. Infect Immun. 1978 Mar;19(3):1097–1098. doi: 10.1128/iai.19.3.1097-1098.1978. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Morris J. A., Stevens A. E., Sojka W. J. Preliminary characterization of cell-free K99 antigen isolated from Escherichia coli B41. J Gen Microbiol. 1977 Apr;99(2):353–357. doi: 10.1099/00221287-99-2-353. [DOI] [PubMed] [Google Scholar]
  16. Morris J. A., Thorns C. J., Sojka W. J. Evidence for two adhesive antigens on the K99 reference strain Escherichia coli B41. J Gen Microbiol. 1980 May;118(1):107–113. doi: 10.1099/00221287-118-1-107. [DOI] [PubMed] [Google Scholar]
  17. Stirm S., Orskov F., Orskov I., Mansa B. Episome-carried surface antigen K88 of Escherichia coli. II. Isolation and chemical analysis. J Bacteriol. 1967 Feb;93(2):731–739. doi: 10.1128/jb.93.2.731-739.1967. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. de Graaf F. K., Klaasen-Boor P., van Hees J. E. Biosynthesis of the K99 surface antigen is repressed by alanine. Infect Immun. 1980 Oct;30(1):125–128. doi: 10.1128/iai.30.1.125-128.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. de Graaf F. K., Wientjes F. B., Klaasen-Boor P. Production of K99 antigen by enterotoxigenic Escherichia coli strains of antigen groups o8, o9, o20, and o101 grown at different conditions. Infect Immun. 1980 Jan;27(1):216–221. doi: 10.1128/iai.27.1.216-221.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. van Embden J. D., de Graaf F. K., Schouls L. M., Teppema J. S. Cloning and expression of a deoxyribonucleic acid fragment that encodes for the adhesive antigen K99. Infect Immun. 1980 Sep;29(3):1125–1133. doi: 10.1128/iai.29.3.1125-1133.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from Infection and Immunity are provided here courtesy of American Society for Microbiology (ASM)

RESOURCES