Translocation, Entry into the Cell

Beard, Matthew

doi:10.1007/978-1-4614-9454-6_7

Cited by 5 publications

(3 citation statements)

References 92 publications

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“…BoNTs enter target cells via a receptor-mediated endocytosis (Beard, 2014;Rossetto, Pirazzini, & Montecucco, 2014), but the precise mechanism of endocytosis according to the cell types is still elusive. BoNT/A has been found to enter neuronal and intestinal cells by a dynamin-dependent pathway in both cell types (Couesnon, Shimizu, & Popoff, 2009).…”

Section: Hcb Enters Neuronal and Intestinal Cells In A Dynamin-depementioning

confidence: 99%

Botulinum neurotoxin type B uses a distinct entry pathway mediated by CDC42 into intestinal cells versus neuronal cells

Connan

Voillequin

Chavez

et al. 2017

Cellular Microbiology

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Botulinum neurotoxins (BoNTs) are responsible for severe flaccid paralysis by inhibiting the release of acetylcholine at the neuromuscular junctions. BoNT type B (BoNT/B) most often induces mild forms of botulism with predominant dysautonomic symptoms. In food borne botulism and botulism by intestinal colonisation such as infant botulism, which are the most frequent naturally acquired forms of botulism, the digestive tract is the main entry route of BoNTs into the organism. We previously showed that BoNT/B translocates through mouse intestinal barrier by an endocytosis-dependent mechanism and subsequently targets neuronal cells, mainly cholinergic neurons, in the intestinal mucosa and musculosa. Here, we investigated the entry pathway of BoNT/B using fluorescent C-terminal domain of the heavy chain (HcB), which is involved in the binding to specific receptor(s) and entry process into target cells. While the combination of gangliosides GD /GD /GT and synaptotagmin I and to a greater extent synaptotagmin II constitutes the functional HcB receptor on NG108-15 neuronal cells, HcB only uses the gangliosides GD /GD /GT to efficiently bind to m-IC intestinal cells. HcB enters both cell types by a dynamin-dependent endocytosis, which is efficiently prevented by Dynasore, a dynamin inhibitor, and reaches a common early endosomal compartment labeled by early endosome antigen (EEA1). In contrast to neuronal cells, HcB uses a Cdc42-dependent pathway to enter intestinal cells. Then, HcB is transported to late endosomes in neuronal cells, whereas it exploits a nonacidified pathway from apical to basal lateral side of m-IC cells supporting a transcytotic route in epithelial intestinal cells.

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Section: Hcb Enters Neuronal and Intestinal Cells In A Dynamin-depementioning

confidence: 99%

Botulinum neurotoxin type B uses a distinct entry pathway mediated by CDC42 into intestinal cells versus neuronal cells

Connan

Voillequin

Chavez

et al. 2017

Cellular Microbiology

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“…The LC is a specific protease that cleaves soluble N -ethyl-maleimide-sensitive factor attachment protein receptor proteins (SNARE proteins) inside target neurons [ 9 ]. The most N-terminal of the HC domains (H n ) contains a transmembrane translocation activity that transports the LC protease into the neuronal cytoplasm [ 10 , 11 ]. The most C-terminal HC domain (H cc ) contains neuron-specific high-affinity binding activity [ 12 , 13 ].…”

Section: Introductionmentioning

confidence: 99%

Purification and Characterization of Recombinant Botulinum Neurotoxin Serotype FA, Also Known as Serotype H

Hackett

Moore

Burgin

et al. 2018

Toxins

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We have purified and characterized recombinant botulinum neurotoxin serotype FA (BoNT/FA). This protein has also been named as a new serotype (serotype H), but the classification has been controversial. A lack of well-characterized, highly pure material has been a roadblock to study. Here we report purification and characterization of enzymatically active, and of inactive nontoxic, recombinant forms of BoNT/FA as tractable alternatives to purifying this neurotoxin from native Clostridium botulinum. BoNT/FA cleaves the same intracellular target proteins as BoNT/F1 and other F serotype BoNTs; the intracellular targets are vesicle associated membrane proteins (VAMP) 1, 2 and 3. BoNT/FA cleaves the same site in VAMP-2 as BoNT/F5, which is different from the cleavage site of other F serotype BoNTs. BoNT/FA has slower enzyme kinetics than BoNT/F1 in a cell-free protease assay and is less potent at inhibiting ex vivo nerve-stimulated skeletal muscle contraction. In contrast, BoNT/FA is more potent at inhibiting neurotransmitter release from cultured neurons.

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“…Following entry into the synaptic vesicle, the proton pumping action of the v-ATPase present on the synaptic vesicle membrane, responsible for the loading of neurotransmitters into the vesicle, will acidify the organelle and produce the necessary environment for the LC to translocate. Treatments that inhibit internalisation, synaptic vesicle recycling, or acidification also inhibit BoNT action [60].…”

Section: Translocation Domainmentioning

confidence: 99%

Botulinum Neurotoxin: A Multifunctional Protein for the Development of New Therapeutics

Fonfría¹

2018

Neurotoxins

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Botulinum neurotoxin (BoNT) is a major therapeutic agent licensed in neurological indications such as dystonia and spasticity. In recent years, its use has steadily increased in other neurological areas and new therapeutic areas and also in the aesthetic setting. Paradoxically, BoNT is also the causative agent of the disease botulism and a potential bioterrorism toxin. The BoNT family of toxins comprised more than 40 individual members, classified into 7 serotypes and are produced by Gram-positive obligate anaerobic bacteria. BoNTs are enzymatic multi-modular proteins with a complex multistep mechanism of action. Their target site is at peripheral neurons, particularly the neuromuscular junction, at which they inhibit acetylcholine neurotransmission. Despite intense activity in the BoNT field, today there are still gaps in knowledge both in clinical practice and in basic research. The discovery of the structure-function of BoNT and its domains has allowed rational design of new features using molecular engineering. The diversity of BoNT molecules, both natural and engineered, is an invaluable pool from which to design future new therapeutics with unique pharmacological properties for current and novel indications.

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Translocation, Entry into the Cell

Cited by 5 publications

References 92 publications

Botulinum neurotoxin type B uses a distinct entry pathway mediated by CDC42 into intestinal cells versus neuronal cells

Botulinum neurotoxin type B uses a distinct entry pathway mediated by CDC42 into intestinal cells versus neuronal cells

Purification and Characterization of Recombinant Botulinum Neurotoxin Serotype FA, Also Known as Serotype H

Botulinum Neurotoxin: A Multifunctional Protein for the Development of New Therapeutics

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