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

Research output: Contribution to journalArticle

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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
Publication statusPublished - 1998
Externally publishedYes

Fingerprint

Clostridium botulinum type B
Abelmoschus
Botulism
Neurotoxins
Japan
Synaptosomes
Binding Sites
Synaptotagmin II
Synaptotagmin I
Vesicle-Associated Membrane Protein 2
Synaptotagmins
Maltose-Binding Proteins
Lipids
Light
Gangliosides
Monoclonal Antibodies
Food
Antibodies

ASJC Scopus subject areas

  • Immunology

Cite this

Kozaki, S., Kamata, Y., Nishiki, T., Kakinuma, H., Maruyama, H., Takahashi, H., ... Nakamura, S. (1998). Characterization of Clostridium botulinum type B neurotoxin associated with infant botulism in Japan. Infection and Immunity, 66(10), 4811-4816.

Characterization of Clostridium botulinum type B neurotoxin associated with infant botulism in Japan. / Kozaki, Shunji; Kamata, Yoichi; Nishiki, Tei-ichi; Kakinuma, Hiroaki; Maruyama, Hiromi; Takahashi, Hiroaki; Karasawa, Tadahiro; Yamakawa, Kiyotaka; Nakamura, Shinichi.

In: Infection and Immunity, Vol. 66, No. 10, 1998, p. 4811-4816.

Research output: Contribution to journalArticle

Kozaki, S, Kamata, Y, Nishiki, T, 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, vol. 66, no. 10, pp. 4811-4816.
Kozaki S, Kamata Y, Nishiki T, Kakinuma H, Maruyama H, Takahashi H et al. Characterization of Clostridium botulinum type B neurotoxin associated with infant botulism in Japan. Infection and Immunity. 1998;66(10):4811-4816.
Kozaki, Shunji ; Kamata, Yoichi ; Nishiki, Tei-ichi ; Kakinuma, Hiroaki ; Maruyama, Hiromi ; Takahashi, Hiroaki ; Karasawa, Tadahiro ; Yamakawa, Kiyotaka ; Nakamura, Shinichi. / Characterization of Clostridium botulinum type B neurotoxin associated with infant botulism in Japan. In: Infection and Immunity. 1998 ; Vol. 66, No. 10. pp. 4811-4816.
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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.",
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