Characterization of Clostridium botulinum type B neurotoxin associated with infant botulism in Japan

Shunji Kozaki, Yoichi Kamata, Tei Ichi Nishiki, Hiroaki Kakinuma, Hiromi Maruyama, Hiroaki Takahashi, Tadahiro Karasawa, Kiyotaka Yamakawa, Shinichi Nakamura

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39 Citations (Scopus)

Abstract

The neurotoxin of strain 111 (111/NT) associated with type B infant botulism showed antigenic and biological properties different from that (Okra/NT) produced by a food-borne botulism-related strain, Okra. The specific toxicity of 111/NT was found to be about 10 times lower than that of Okra/NT. The monoclonal antibodies recognizing the light chain cross-reacted with both neurotoxins, whereas most of the antibodies recognizing the carboxyl-terminal half of the heavy chain of Okra/NT did not react to 111/ONT. Binding experiments with rat brain synaptosomes revealed that 125I-labeled 111/NT bound to a single binding site with a dissociation constant (K(d)) of 2.5 nM; the value was rather lower than that (0.42 nM) of 125I-Okra/NT for the high-affinity binding site. In the lipid vesicles reconstituted with ganglioside GTlb, 125I-Okra/NT interacted with the amino-terminal domain of synaptotagmin 1 (Stg1N) or synaptotagmin 2 (Stg2N), fused with the maltose-binding protein, in the same manner as the respective full-length synaptotagmins, and the K(d) values accorded with those of the low- and high-affinity binding sites in synaptosomes. However, 125I- 111/NT only exhibited a low capacity for binding to the lipid vesicles containing Stg2N, but not Stg1N, in the presence of ganglioside GT1b. Moreover, synaptobrevin-2, an intracellular target protein, was digested to the same extent by the light chains of both neurotoxins in a concentration- dependent manner. These findings indicate that the 111/NT molecule possesses the receptor-recognition site structurally different from Okra/NT, probably causing a decreased specific toxicity.

Original languageEnglish
Pages (from-to)4811-4816
Number of pages6
JournalInfection and Immunity
Volume66
Issue number10
DOIs
Publication statusPublished - 1998

ASJC Scopus subject areas

  • Parasitology
  • Microbiology
  • Immunology
  • Infectious Diseases

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    Kozaki, S., Kamata, Y., Nishiki, T. I., Kakinuma, H., Maruyama, H., Takahashi, H., Karasawa, T., Yamakawa, K., & Nakamura, S. (1998). Characterization of Clostridium botulinum type B neurotoxin associated with infant botulism in Japan. Infection and Immunity, 66(10), 4811-4816. https://doi.org/10.1128/iai.66.10.4811-4816.1998