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. 1987 May;55(5):1294–1299. doi: 10.1128/iai.55.5.1294-1299.1987

Maintenance of biological activity of pertussis toxin radioiodinated while bound to fetuin-agarose.

G D Armstrong, M S Peppler
PMCID: PMC260504  PMID: 2437034

Abstract

We developed a method to produce radioiodinated pertussis toxin (PT) which was active in the goose erythrocyte agglutination and CHO cell assay systems. The procedure used fetuin coupled to agarose to prevent inactivation of the toxin during the iodination reaction. Analysis of the labeled PT by affinity chromatography on fetuin-agarose and wheat germ agglutinin-agarose and by sodium dodecyl sulfate-polyacrylamide gel electrophoresis indicated that there were minimal amounts of labeled fetuin or other contaminants in the labeled PT preparations. All five of the subunits of the toxin appeared to be labeled by the procedure. The labeling method will facilitate further investigations into the nature of the interaction and activity of PT in host tissues.

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Selected References

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  1. Arai H., Sato Y. Separation and characterization of two distinct hemagglutinins contained in purified leukocytosis-promoting factor from Bordetella pertussis. Biochim Biophys Acta. 1976 Oct 22;444(3):765–782. doi: 10.1016/0304-4165(76)90323-8. [DOI] [PubMed] [Google Scholar]
  2. Burns D. L., Kenimer J. G., Manclark C. R. Role of the A subunit of pertussis toxin in alteration of Chinese hamster ovary cell morphology. Infect Immun. 1987 Jan;55(1):24–28. doi: 10.1128/iai.55.1.24-28.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Cuatrecasas P. Interaction of Vibrio cholerae enterotoxin with cell membranes. Biochemistry. 1973 Aug 28;12(18):3547–3558. doi: 10.1021/bi00742a031. [DOI] [PubMed] [Google Scholar]
  4. Cuatrecasas P., Parikh I., Hollenberg M. D. Affinity chromatography and structural analysis of Vibrio cholerae enterotoxin-ganglioside agarose and the biological effects of ganglioside-containing soluble polymers. Biochemistry. 1973 Oct 9;12(21):4253–4264. doi: 10.1021/bi00745a033. [DOI] [PubMed] [Google Scholar]
  5. Endoh M., Fujimoto T., Nakase Y. Distribution of radioactivity in rats injected intravenously with 125I-labeled histamine-sensitizing factor of Bordetella pertussis. Microbiol Immunol. 1984;28(2):233–236. doi: 10.1111/j.1348-0421.1984.tb00674.x. [DOI] [PubMed] [Google Scholar]
  6. Fishman P. H. Role of membrane gangliosides in the binding and action of bacterial toxins. J Membr Biol. 1982;69(2):85–97. doi: 10.1007/BF01872268. [DOI] [PubMed] [Google Scholar]
  7. Hewlett E. L., Sauer K. T., Myers G. A., Cowell J. L., Guerrant R. L. Induction of a novel morphological response in Chinese hamster ovary cells by pertussis toxin. Infect Immun. 1983 Jun;40(3):1198–1203. doi: 10.1128/iai.40.3.1198-1203.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Irons L. I., MacLennan A. P. Isolation of the lymphocytosis promoting factor-haemagglutinin of Bordetella pertussis by affinity chromatography. Biochim Biophys Acta. 1979 Sep 29;580(1):175–185. doi: 10.1016/0005-2795(79)90208-3. [DOI] [PubMed] [Google Scholar]
  9. Katada T., Tamura M., Ui M. The A protomer of islet-activating protein, pertussis toxin, as an active peptide catalyzing ADP-ribosylation of a membrane protein. Arch Biochem Biophys. 1983 Jul 1;224(1):290–298. doi: 10.1016/0003-9861(83)90212-6. [DOI] [PubMed] [Google Scholar]
  10. Locht C., Keith J. M. Pertussis toxin gene: nucleotide sequence and genetic organization. Science. 1986 Jun 6;232(4755):1258–1264. doi: 10.1126/science.3704651. [DOI] [PubMed] [Google Scholar]
  11. Manning D. R., Fraser B. A., Kahn R. A., Gilman A. G. ADP-ribosylation of transducin by islet-activation protein. Identification of asparagine as the site of ADP-ribosylation. J Biol Chem. 1984 Jan 25;259(2):749–756. [PubMed] [Google Scholar]
  12. Miller D. L., Ross E. M., Alderslade R., Bellman M. H., Rawson N. S. Pertussis immunisation and serious acute neurological illness in children. Br Med J (Clin Res Ed) 1981 May 16;282(6276):1595–1599. doi: 10.1136/bmj.282.6276.1595. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Milligan G., Gierschik P., Spiegel A. M., Klee W. A. The GTP-binding regulatory proteins of neuroblastoma x glioma, NG108-15, and glioma, C6, cells. Immunochemical evidence of a pertussis toxin substrate that is neither Ni nor No. FEBS Lett. 1986 Jan 20;195(1-2):225–230. doi: 10.1016/0014-5793(86)80165-x. [DOI] [PubMed] [Google Scholar]
  14. Munoz J. J., Arai H., Cole R. L. Mouse-protecting and histamine-sensitizing activities of pertussigen and fimbrial hemagglutinin from Bordetella pertussis. Infect Immun. 1981 Apr;32(1):243–250. doi: 10.1128/iai.32.1.243-250.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Nicolson G. L., Irimura T. Estimating glycoprotein carbohydrate chain structures by lectin reactivities in polyacrylamide gels. Biol Cell. 1984;51(2):157–164. doi: 10.1111/j.1768-322x.1984.tb00294.x. [DOI] [PubMed] [Google Scholar]
  16. Nicosia A., Perugini M., Franzini C., Casagli M. C., Borri M. G., Antoni G., Almoni M., Neri P., Ratti G., Rappuoli R. Cloning and sequencing of the pertussis toxin genes: operon structure and gene duplication. Proc Natl Acad Sci U S A. 1986 Jul;83(13):4631–4635. doi: 10.1073/pnas.83.13.4631. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Nogimori K., Ito K., Tamura M., Satoh S., Ishii S., Ui M. Chemical modification of islet-activating protein, pertussis toxin. Essential role of free amino groups in its lymphocytosis-promoting activity. Biochim Biophys Acta. 1984 Sep 28;801(2):220–231. doi: 10.1016/0304-4165(84)90071-0. [DOI] [PubMed] [Google Scholar]
  18. Nogimori K., Tamura M., Yajima M., Hashimura N., Ishii S., Ui M. Structure-function relationship of islet-activating protein, pertussis toxin: biological activities of hybrid toxins reconstituted from native and methylated subunits. Biochemistry. 1986 Mar 25;25(6):1355–1363. doi: 10.1021/bi00354a025. [DOI] [PubMed] [Google Scholar]
  19. Nogimori K., Tamura M., Yajima M., Ito K., Nakamura T., Kajikawa N., Maruyama Y., Ui M. Dual mechanisms involved in development of diverse biological activities of islet-activating protein, pertussis toxin, as revealed by chemical modification of lysine residues in the toxin molecule. Biochim Biophys Acta. 1984 Sep 28;801(2):232–243. doi: 10.1016/0304-4165(84)90072-2. [DOI] [PubMed] [Google Scholar]
  20. Peppler M. S., Judd R. C., Munoz J. J. Effect of proteolytic enzymes, storage and reduction on the structure and biological activity of pertussigen, a toxin from Bordetella pertussis. Dev Biol Stand. 1985;61:75–87. [PubMed] [Google Scholar]
  21. Sato H., Sato Y. Bordetella pertussis infection in mice: correlation of specific antibodies against two antigens, pertussis toxin, and filamentous hemagglutinin with mouse protectivity in an intracerebral or aerosol challenge system. Infect Immun. 1984 Nov;46(2):415–421. doi: 10.1128/iai.46.2.415-421.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Sekura R. D., Fish F., Manclark C. R., Meade B., Zhang Y. L. Pertussis toxin. Affinity purification of a new ADP-ribosyltransferase. J Biol Chem. 1983 Dec 10;258(23):14647–14651. [PubMed] [Google Scholar]
  23. Spiro R. G. Studies on fetuin, a glycoprotein of fetal serum. I. Isolation, chemical composition, and physiochemical properties. J Biol Chem. 1960 Oct;235(10):2860–2869. [PubMed] [Google Scholar]
  24. Spivak J. L., Small D., Hollenberg M. D. Erythropoietin: isolation by affinity chromatography with lectin-agarose derivatives. Proc Natl Acad Sci U S A. 1977 Oct;74(10):4633–4635. doi: 10.1073/pnas.74.10.4633. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Steinman L., Weiss A., Adelman N., Lim M., Zuniga R., Oehlert J., Hewlett E., Falkow S. Pertussis toxin is required for pertussis vaccine encephalopathy. Proc Natl Acad Sci U S A. 1985 Dec;82(24):8733–8736. doi: 10.1073/pnas.82.24.8733. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Sternweis P. C., Robishaw J. D. Isolation of two proteins with high affinity for guanine nucleotides from membranes of bovine brain. J Biol Chem. 1984 Nov 25;259(22):13806–13813. [PubMed] [Google Scholar]
  27. Takasaki S., Kobata A. Asparagine-linked sugar chains of fetuin: occurrence of tetrasialyl triantennary sugar chains containing the Gal beta 1----3GlcNAc sequence. Biochemistry. 1986 Sep 23;25(19):5709–5715. doi: 10.1021/bi00367a054. [DOI] [PubMed] [Google Scholar]
  28. Tamura M., Nogimori K., Murai S., Yajima M., Ito K., Katada T., Ui M., Ishii S. Subunit structure of islet-activating protein, pertussis toxin, in conformity with the A-B model. Biochemistry. 1982 Oct 26;21(22):5516–5522. doi: 10.1021/bi00265a021. [DOI] [PubMed] [Google Scholar]
  29. Tamura M., Nogimori K., Yajima M., Ase K., Ui M. A role of the B-oligomer moiety of islet-activating protein, pertussis toxin, in development of the biological effects on intact cells. J Biol Chem. 1983 Jun 10;258(11):6756–6761. [PubMed] [Google Scholar]
  30. Van Dop C., Yamanaka G., Steinberg F., Sekura R. D., Manclark C. R., Stryer L., Bourne H. R. ADP-ribosylation of transducin by pertussis toxin blocks the light-stimulated hydrolysis of GTP and cGMP in retinal photoreceptors. J Biol Chem. 1984 Jan 10;259(1):23–26. [PubMed] [Google Scholar]

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