Binding of Clostridium botulinum type C and D neurotoxins to ganglioside and phospholipid

Novel insights into the receptor for clostridial neurotoxins

Kentaro Tsukamoto, Tomoko Kohda, Masafumi Mukamoto, Kumiko Takeuchi, Hideshi Ihara, Masaki Saito, Shunji Kozaki

Research output: Contribution to journalArticle

101 Citations (Scopus)

Abstract

Clostridium botulinum neurotoxins (BoNTs) act on nerve endings to block acetylcholine release. Their potency is due to their enzymatic activity and selective high affinity binding to neurons. Although there are many pieces of data available on the receptor for BoNT, little attempt has been made to characterize the receptors for BoNT/C and BoNT/D. For this purpose, we prepared the recombinant carboxyl-terminal domain of the heavy chain (HC) and then examined its binding capability to rat brain synaptosomes treated with enzymes and heating. Synaptosomes treated with proteinase K or heating retained binding capability to both HC/C and HC/D, suggesting that a proteinaceous substance does not constitute the receptor component. We next performed a thin layer chromatography overlay assay of HC with a lipid extract of synaptosomes. Under physiological or higher ionic strengths, HC/C bound to gangliosides GD1b and GT1b. These data are in accord with results showing that neuraminidase and endoglycoceramidase treatment decreased HC/C binding to synaptosomes. On the other hand, H C/D interacted with phosphatidylethanolamine but not with any ganglioside. Using cerebellar granule cells obtained from GM3 synthase knock-out mice, we found that BoNT/C did not elicit a toxic effect but that BoNT/D still inhibited glutamate release to the same extent as in granule cells from wild type mice. These observations suggested that BoNT/C recognized GD1b and GT1b as functional receptors, whereas BoNT/D induced toxicity in a ganglioside- independent manner, possibly through binding to phosphatidylethanolamine. Our results provide novel insights into the receptor for clostridial neurotoxin.

Original languageEnglish
Pages (from-to)35164-35171
Number of pages8
JournalJournal of Biological Chemistry
Volume280
Issue number42
DOIs
Publication statusPublished - 21-10-2005

Fingerprint

Clostridium botulinum type D
Clostridium botulinum type C
Tetanus Toxin
Clostridium
Synaptosomes
Gangliosides
Neurotoxins
Phospholipids
endoglycoceramidase
Heating
Thin layer chromatography
Endopeptidase K
Poisons
Neuraminidase
Clostridium botulinum
Ionic strength
Acetylcholine
Neurons
Nerve Endings
Toxicity

All Science Journal Classification (ASJC) codes

  • Biochemistry
  • Molecular Biology
  • Cell Biology

Cite this

Tsukamoto, Kentaro ; Kohda, Tomoko ; Mukamoto, Masafumi ; Takeuchi, Kumiko ; Ihara, Hideshi ; Saito, Masaki ; Kozaki, Shunji. / Binding of Clostridium botulinum type C and D neurotoxins to ganglioside and phospholipid : Novel insights into the receptor for clostridial neurotoxins. In: Journal of Biological Chemistry. 2005 ; Vol. 280, No. 42. pp. 35164-35171.
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Binding of Clostridium botulinum type C and D neurotoxins to ganglioside and phospholipid : Novel insights into the receptor for clostridial neurotoxins. / Tsukamoto, Kentaro; Kohda, Tomoko; Mukamoto, Masafumi; Takeuchi, Kumiko; Ihara, Hideshi; Saito, Masaki; Kozaki, Shunji.

In: Journal of Biological Chemistry, Vol. 280, No. 42, 21.10.2005, p. 35164-35171.

Research output: Contribution to journalArticle

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