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Infect Immun. Nov 1989; 57(11): 3324–3330.
PMCID: PMC259811

Binding of pertussis toxin to eucaryotic cells and glycoproteins.

Abstract

The binding of pertussis toxin and its subunits to cell surface receptors and purified glycoproteins was examined. The interaction of pertussis toxin with components of two variant Chinese hamster ovary (CHO) cell lines was studied. These cell lines are deficient in either sialic acid residues (LEC 2) or sialic acid and galactose residues (LEC 8) on cell surface macromolecules. The binding of pertussis toxin to components of these cells differed from the binding of the toxin to wild-type components. Although the toxin bound to a 165,000-dalton glycoprotein found in N-octylglucoside extracts of wild-type cells, it did not bind to components found in extracts of LEC 2 cells. In contrast, the toxin bound to components found in extracts of LEC 8 cells, which are variant cells that contain increased amounts of terminal N-acetylglucosamine residues on cell surface macromolecules. These results suggest that the receptor for pertussis toxin on CHO cells contains terminal acetamido-containing sugars. The cytopathic effect of the toxin on both types of variant cells was much reduced compared with its effects on wild-type cells. Thus, optimal functional binding of pertussis toxin appears to require a complete sialyllactosamine (NeuAc----Gal beta 4GlcNAc) sequence on surface macromolecules. In addition to studying the nature of the eucaryotic receptor for pertussis toxin, we examined corresponding binding sites for glycoproteins on the toxin molecule. Binding of both S2-S4 and S3-S4 dimers of the toxin to cellular components and purified glycoproteins was observed. The two dimers bound to a number of glycoproteins containing N-linked oligosaccharides but not O-linked oligosaccharides, and differences in the binding of the two dimers to some glycoproteins was noted. These data indicate that the holotoxin molecule contains at least two glycoprotein-binding sites which may have slightly different specificities for glycoproteins.

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  • Armstrong GD, Howard LA, Peppler MS. Use of glycosyltransferases to restore pertussis toxin receptor activity to asialoagalactofetuin. J Biol Chem. 1988 Jun 25;263(18):8677–8684. [PubMed]
  • Bradford MM. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem. 1976 May 7;72:248–254. [PubMed]
  • Brennan MJ, David JL, Kenimer JG, Manclark CR. Lectin-like binding of pertussis toxin to a 165-kilodalton Chinese hamster ovary cell glycoprotein. J Biol Chem. 1988 Apr 5;263(10):4895–4899. [PubMed]
  • Briles EB. Lectin-resistant cell surface variants of eukaryotic cells. Int Rev Cytol. 1982;75:101–165. [PubMed]
  • Burnette WN. "Western blotting": electrophoretic transfer of proteins from sodium dodecyl sulfate--polyacrylamide gels to unmodified nitrocellulose and radiographic detection with antibody and radioiodinated protein A. Anal Biochem. 1981 Apr;112(2):195–203. [PubMed]
  • Burns DL, Hewlett EL, Moss J, Vaughan M. Pertussis toxin inhibits enkephalin stimulation of GTPase of NG108-15 cells. J Biol Chem. 1983 Feb 10;258(3):1435–1438. [PubMed]
  • Burns DL, Kenimer JG, Manclark CR. Role of the A subunit of pertussis toxin in alteration of Chinese hamster ovary cell morphology. Infect Immun. 1987 Jan;55(1):24–28. [PMC free article] [PubMed]
  • Capiau C, Petre J, Van Damme J, Puype M, Vandekerckhove J. Protein-chemical analysis of pertussis toxin reveals homology between the subunits S2 and S3, between S1 and the A chains of enterotoxins of Vibrio cholerae and Escherichia coli and identifies S2 as the haptoglobin-binding subunit. FEBS Lett. 1986 Aug 18;204(2):336–340. [PubMed]
  • Casey PJ, Gilman AG. G protein involvement in receptor-effector coupling. J Biol Chem. 1988 Feb 25;263(6):2577–2580. [PubMed]
  • Deutscher SL, Hirschberg CB. Mechanism of galactosylation in the Golgi apparatus. A Chinese hamster ovary cell mutant deficient in translocation of UDP-galactose across Golgi vesicle membranes. J Biol Chem. 1986 Jan 5;261(1):96–100. [PubMed]
  • Deutscher SL, Nuwayhid N, Stanley P, Briles EI, Hirschberg CB. Translocation across Golgi vesicle membranes: a CHO glycosylation mutant deficient in CMP-sialic acid transport. Cell. 1984 Dec;39(2 Pt 1):295–299. [PubMed]
  • Dzandu JK, Deh ME, Barratt DL, Wise GE. Detection of erythrocyte membrane proteins, sialoglycoproteins, and lipids in the same polyacrylamide gel using a double-staining technique. Proc Natl Acad Sci U S A. 1984 Mar;81(6):1733–1737. [PMC free article] [PubMed]
  • Furthmayr H, Marchesi VT. Glycophorins: isolation, orientation, and localization of specific domains. Methods Enzymol. 1983;96:268–280. [PubMed]
  • Gilman AG. G proteins and dual control of adenylate cyclase. Cell. 1984 Mar;36(3):577–579. [PubMed]
  • Gilman AG. G proteins: transducers of receptor-generated signals. Annu Rev Biochem. 1987;56:615–649. [PubMed]
  • Hewlett EL, Sauer KT, Myers GA, Cowell JL, Guerrant RL. Induction of a novel morphological response in Chinese hamster ovary cells by pertussis toxin. Infect Immun. 1983 Jun;40(3):1198–1203. [PMC free article] [PubMed]
  • Katada T, Ui M. Direct modification of the membrane adenylate cyclase system by islet-activating protein due to ADP-ribosylation of a membrane protein. Proc Natl Acad Sci U S A. 1982 May;79(10):3129–3133. [PMC free article] [PubMed]
  • Katada T, Ui M. ADP ribosylation of the specific membrane protein of C6 cells by islet-activating protein associated with modification of adenylate cyclase activity. J Biol Chem. 1982 Jun 25;257(12):7210–7216. [PubMed]
  • Li E, Becker A, Stanley SL., Jr Use of Chinese hamster ovary cells with altered glycosylation patterns to define the carbohydrate specificity of Entamoeba histolytica adhesion. J Exp Med. 1988 May 1;167(5):1725–1730. [PMC free article] [PubMed]
  • Locht C, Keith JM. Pertussis toxin gene: nucleotide sequence and genetic organization. Science. 1986 Jun 6;232(4755):1258–1264. [PubMed]
  • Martin GR, Timpl R. Laminin and other basement membrane components. Annu Rev Cell Biol. 1987;3:57–85. [PubMed]
  • Meade BD, Kind PD, Manclark CR. Lymphocytosis-promoting factor of Bordetella pertussis alters mononuclear phagocyte circulation and response to inflammation. Infect Immun. 1984 Dec;46(3):733–739. [PMC free article] [PubMed]
  • Morse SI, Morse JH. Isolation and properties of the leukocytosis- and lymphocytosis-promoting factor of Bordetella pertussis. J Exp Med. 1976 Jun 1;143(6):1483–1502. [PMC free article] [PubMed]
  • Nakane PK, Kawaoi A. Peroxidase-labeled antibody. A new method of conjugation. J Histochem Cytochem. 1974 Dec;22(12):1084–1091. [PubMed]
  • Nicosia A, Perugini M, Franzini C, Casagli MC, Borri MG, 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. [PMC free article] [PubMed]
  • 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. [PubMed]
  • 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. [PubMed]
  • 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]
  • Yajima M, Hosoda K, Kanbayashi Y, Nakamura T, Takahashi I, Ui M. Biological properties of islets-activating protein (IAP) purified from the culture medium of Bordetella pertussis. J Biochem. 1978 Jan;83(1):305–312. [PubMed]

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