Hemagglutinin‐33 of type A botulinum neurotoxin complex binds with synaptotagmin II

Zhou, Yu; Foss, Sean Erik; Lindo, Paul; Sarkar, Hemanta K.; Singh, Bal Ram

doi:10.1111/j.1742-4658.2005.04688.x

Cited by 17 publications

(10 citation statements)

References 40 publications

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“…Previous protection and transmission electron microscopy studies have shown detailed interactions of BoNT/D NTNH and HA70 (Hasegawa et al, 2007; Niwa et al, 2007); HA33 was shown to bind epithelial cells, possibly bringing the toxin in close proximity to its receptors (Matsumura et al, 2008; Zhou et al, 2005). Perhaps the larger BoNT/B complex, with presumably more NAP proteins, can bind intestinal membranes with a higher affinity than the BoNT/A complexes.…”

Section: Discussionmentioning

confidence: 98%

Comparison of oral toxicological properties of botulinum neurotoxin serotypes A and B

Cheng

Henderson

2011

Toxicon

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Botulinum neurotoxins (BoNTs) are among the most potent biological toxins for humans. Of the seven known serotypes (A-G) of BoNT, serotypes A, B and E cause most of the foodborne intoxications in humans. BoNTs in nature are associated with non-toxic accessory proteins known as neurotoxin-associated proteins (NAPs), forming large complexes that have been shown to play important roles in oral toxicity. Using mouse intraperitoneal and oral models of botulism, we determined the dose response to both BoNT/B holotoxin and complex toxins, and compared the toxicities of BoNT/B and BoNT/A complexes. Although serotype A and B complexes have similar NAP composition, BoNT/B formed larger-sized complexes, and was approximately 90 times more lethal in mouse oral intoxications than BoNT/A complexes. When normalized by mean lethal dose, mice orally treated with high doses of BoNT/B complex showed a delayed time-to-death when compared with mice treated with BoNT/A complex. Furthermore, we determined the effect of various food matrices on oral toxicity of BoNT/A and BoNT/B complexes. BoNT/B complexes showed lower oral bioavailability in liquid egg matrices when compared to BoNT/A complexes. In summary, our studies revealed several factors that can either enhance or reduce the toxicity and oral bioavailability of BoNTs. Dissecting the complexities of the different BoNT serotypes and their roles in foodborne botulism will lead to a better understanding of toxin biology and aid future food risk assessments.

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Section: Discussionmentioning

confidence: 98%

Comparison of oral toxicological properties of botulinum neurotoxin serotypes A and B

Cheng

Henderson

2011

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“…HA‐33/HA‐17 arms facilitated cell binding in addition to transport through the cell layer. Zhou et al . (2005) isolated synaptotagmin II using a pull‐down assay from the rat brain synaptosomal protein extract as a putative protein receptor for HA‐33 on nerve cells.…”

Section: Discussionmentioning

confidence: 99%

HA-33 facilitates transport of the serotype D botulinum toxin across a rat intestinal epithelial cell monolayer

Ito

Sagane

Miyata

et al. 2011

FEMS Immunol Med Microbiol

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A large size botulinum toxin complex (L-TC) is composed of a single neurotoxin (BoNT), a single nontoxic nonhaemagglutinin (NTNHA) and a haemagglutinin (HA) complex. The HA complex is comprised of three HA-70 molecules and three arm structures of HA-33/HA-17 that consist of two HA-33 and a single HA-17. In addition to the mature L-TC, smaller TCs are present in cultures: M-TC (BoNT/NTNHA), M-TC/HA-70 and immature L-TCs with fewer HA-33/HA-17 arms than mature L-TC. Because L-TC displays higher oral toxicity than pure BoNT, it was presumed that nontoxic proteins are critical for food poisoning. In this study, the absorption of TCs across intestinal epithelial cells was assessed by examining the cell binding and monolayer transport of serotype D toxins in the rat intestinal epithelial cell line IEC-6. All TCs, including pure BoNT, displayed binding and transport, with mature L-TC showing the greatest potency. Inhibition experiments using antibodies revealed that BoNT, HA-70 and HA-33 could be responsible for the binding and transport. The findings here indicate that all TCs can transport across the cell layer via a sialic acid-dependent process. Nonetheless, binding and transport markedly increased with number of HA-33/HA-17 arms in the TC. We therefore conclude that the HA-33/HA-17 arm is not necessarily required for, but facilitates, transport of botulinum toxin complexes.

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“…44 A reasonable explanation for this observation is that the BoNT/A in the sample, was partially in complex form containing hemagglutinin-33, which is a neurotoxin-associated protein that can bind synaptotagmin II and trigger cellular responses. 45 This suggests that the p21-Fc has the potential to detect both of BoNT/A complex in addition to BoNT/B. But the cell sensor presensitized with p21-Fc was limit to distinguish these two type of BoNTs.…”

Section: Detection Of Bont/b By the Mast Cell Biosensormentioning

confidence: 99%

Development of Aequorin-Based Mast Cell Nanosensor for Rapid Identification of Botulinum Neurotoxin Type B

Xin¹,

Yao²,

Gao³

et al. 2014

j biomed nanotechnol

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An aequorin-based mast cell sensor was developed for rapid identification and detection of protein toxins. To monitor mast cell activation and to improve the sensitivity of detection, aequorin was stably expressed in the cells and used as an indicator of calcium flux and the stable cell line was created. The procedures for preparing cells were optimized to improve the quantum yield and sensitivity of detection. The cells sensitized with anti-DNP (dinitrophenyl) IgE were capable of detecting as little as 0.1 ng/mL DNP-BSA in less than 2 min. To demonstrate the utility of this mast cell sensor in detecting protein toxins, an IgE chimeric protein (p21-Fc was created. This was achieved by fusing the Fc region of IgE antibody to a 21-amino acid peptide (p21) derived from residues 40-60 of synaptotagmin II (Syt II), the protein receptor of botulinum neurotoxin type B (BoNT/B). In addition, magnetic beads linked with anti-BoNT/B polyclonal antibody were prepared to capture BoNT/B molecules in solution to make targets multivalent and concentrated. The p21-Fc binds to Fc RI receptors on RBL (Rat basophilic leukemia) cells and can be cross-linked by BoNT/B captured by the magnetic beads. This led to cell activation and Ca 2+ flux indicated by an increase of quantum yield. This assay can detect as little as 100 pM (15 ng/mL) BoNT/B in less than 1 hr, which included the initial sample preparation time. This study lays a foundation for detecting other protein toxins using mast cell sensors.

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Hemagglutinin‐33 of type A botulinum neurotoxin complex binds with synaptotagmin II

Cited by 17 publications

References 40 publications

Comparison of oral toxicological properties of botulinum neurotoxin serotypes A and B

Comparison of oral toxicological properties of botulinum neurotoxin serotypes A and B

HA-33 facilitates transport of the serotype D botulinum toxin across a rat intestinal epithelial cell monolayer

Development of Aequorin-Based Mast Cell Nanosensor for Rapid Identification of Botulinum Neurotoxin Type B

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