Characterization of haemagglutinin activity of Clostridium botulinum type C and D 16S toxins, and one subcomponent of haemagglutinin (HA1)

Kaoru Inoue, Yukako Fujinaga, Koichi Honke, Kenji Yokota, Tetsuya Ikeda, Tohru Ohyama, Kouichi Takeshi, Toshihiro Watanabe, Katsuhiro Inoue, Keiji Oguma

Research output: Contribution to journalArticle

49 Citations (Scopus)

Abstract

The 16S toxin and one subcomponent of haemagglutinin (HA), designated HA1, were purified from a type D culture of Clostridium botulinum by a newly established procedure, and their HA activities as well as that of purified type C 16S toxin were characterized. SDS-PAGE analysis indicated that the free HA1 forms a polymer with a molecular mass of approximately 200 kDa. Type C and D 16S toxins agglutinated human erythrocytes in the same manner. Their HA titres were dramatically reduced by employing erythrocytes that had been previously treated with neuraminidase, papain or proteinase K, and were inhibited by the addition of N-acetylneuraminic acid to the reaction mixtures. In a direct-binding test to glycolipids such as SPG (NeuAcα2-Galβ1-4GlcNAcβ1-3Galβ1-4Glcβ1-Cer) and GM3 (NeuAcα2-3Galβ1-4Glcβ1-Cer), and glycoproteins such as glycophorin A and/or B prepared from the erythrocytes, both toxins bound to sialylglycolipids and sialoglycoproteins, but bound to neither neutral glycolipids nor asialoglycoproteins. On the basis of these results, it was concluded that type C and D 16S toxins bind to erythrocytes through N-acetylneuraminic acid. HA1 showed no haemagglutination activity, although it did bind to sialylglycolipids. We therefore speculate that binding to glycoproteins rather than to glycolipids may be important in causing haemagglutination by type C and D 16S toxins.

Original languageEnglish
Pages (from-to)2533-2542
Number of pages10
JournalMicrobiology
Volume145
Issue number9
DOIs
Publication statusPublished - Sep 1999

Keywords

  • Clostridium botulinum
  • Glycolipid
  • Glycoprotein
  • Haemagglutinin

ASJC Scopus subject areas

  • Microbiology

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