Different response of the knockout mice lacking b-series gangliosides against botulinum and tetanus toxins

Kitamura, Masaru; Igimi, Shizunobu; Furukawa, Koichi

doi:10.1016/j.bbadis.2005.04.005

Cited by 38 publications

(25 citation statements)

References 12 publications

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“…Studies in mice with a disruption of the ␣-2,8-sialyltransferase gene (GD3 synthase) displayed an ϳ60-fold decrease in sensitivity to TeNT relative to wild type littermates (30), highlighting the key role of b-series gangliosides in TeNT toxicity. Furthermore, mice lacking all complex gangliosides through disruption of the ␤1,4-N-acetylgalactosaminyltransferase gene (GM2/GD2 synthase) displayed an ϳ600-fold decrease in sensitivity to TeNT relative to wild type littermates.…”

Section: Discussionmentioning

confidence: 99%

Gangliosides as High Affinity Receptors for Tetanus Neurotoxin

Chen

Kim

et al. 2009

Journal of Biological Chemistry

118

134

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Tetanus neurotoxin (TeNT) is an exotoxin produced byClostridium tetani that causes paralytic death to hundreds of thousands of humans annually. TeNT cleaves vesicle-associated membrane protein-2, which inhibits neurotransmitter release in the central nervous system to elicit spastic paralysis, but the molecular basis for TeNT entry into neurons remains unclear. TeNT is a ϳ150-kDa protein that has AB structure-function properties; the A domain is a zinc metalloprotease, and the B domain encodes a translocation domain and C-terminal receptor-binding domain (HCR/T). Earlier studies showed that HCR/T bound gangliosides via two carbohydrate-binding sites, termed the lactose-binding site (the "W" pocket) and the sialic acid-binding site (the "R" pocket). Here we report that TeNT high affinity binding to neurons is mediated solely by gangliosides. Glycan array and solid phase binding analyses identified gangliosides that bound exclusively to either the W pocket or the R pocket of TeNT; GM1a bound to the W pocket, and GD3 bound to the R pocket. Using these gangliosides and mutated forms of HCR/T that lacked one or both carbohydrate-binding pocket, gangliosides binding to both of the W and R pockets were shown to be necessary for high affinity binding to neuronal and non-neuronal cells. The crystal structure of a ternary complex of HCR/T with sugar components of two gangliosides bound to the W and R supported the binding of gangliosides to both carbohydrate pockets. These data show that gangliosides are functional dual receptors for TeNT.Tetanus is an acute, often fatal disease of humans that was first described by Hippocrates over 24 centuries ago (1). Tetanus is characterized by generalized increased rigidity and convulsive spasms of skeletal muscles. Tetanus is caused by exposure to tetanus neurotoxin (TeNT) 3 produced by the sporeforming bacterium Clostridium tetani. TeNT is delivered from the bloodstream to the peripheral nervous system, from where TeNT traffics to the central nervous system to cleave vesicleassociated membrane protein-2 (VAMP2), which inhibits neurotransmitter release and elicits spastic paralysis (2). Although prevented by vaccination, tetanus is responsible for hundreds of thousands of deaths per year in countries where vaccination is not common (3).TeNT is produced as a ϳ150-kDa protein that is cleaved to a di-chain protein, comprising an N-terminal light chain (ϳ50 kDa) and a C-terminal heavy chain domain (ϳ100 kDa) linked through a single disulfide bond (4). TeNT light chain is a zinc metalloprotease that cleaves the neuronal SNARE protein VAMP2 (2). The TeNT heavy chain contains two functional domains: a translocation domain and a C-terminal receptorbinding domain (HCR/T, ϳ50 kDa).The first step in TeNT action involves binding to a receptor(s) on the presynaptic membrane of ␣-motor neurons. Although the molecular basis for TeNT entry remains undetermined, an unambiguous role for gangliosides has been demonstrated (5-9). Current models implicate a dual receptor mechanism for the binding of t...

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Section: Discussionmentioning

confidence: 99%

Gangliosides as High Affinity Receptors for Tetanus Neurotoxin

Chen

Kim

et al. 2009

Journal of Biological Chemistry

118

134

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“…Other notable examples of Siadependent toxins are those from Clostridium botulinum and Clostridium tetani, the causative agents of botulism and tetanus, respectively, which both recognize gangliosides [82].…”

Section: Toxinsmentioning

confidence: 99%

Sialic acid-specific lectins: occurrence, specificity and function

Lehmann

Tiralongo

2006

Cell. Mol. Life Sci.

206

197

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Sialic acids consist of a family of acidic nine-carbon sugars that are typically located at the terminal positions of a variety of glycoconjugates. Naturally occurring sialic acids show an immense diversity of structure, and this reflects their involvement in a variety of biologically important processes. One such process involves the direct participation of sialic acids in recognition events through specific interactions with lectins, a family of proteins that recognise and bind sugars. This review will present a detailed overview of our current knowledge regarding the occurrence, specificity and function of sialic acid-specific lectins, particularly those that occur in viruses, bacteria and non-vertebrate eukaryotes.

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“…TeNT prefers to bind b-series gangliosides (GD1b, GT1b, and GQ1b) (27), and in solid-phase assays, HCR/T binds most avidly to GT1b, GD1b, and GD1a (23). Knockout mice that do not produce b-series gangliosides or complex gangliosides are resistant to TeNT (28,29). To test the ganglioside binding potential of TeNT(RY) relative to HCR/T, Neuro-2a cells (ATCC CCL-131) were loaded with exogenous gangliosides.…”

Section: Properties Of Tent(ry)mentioning

confidence: 99%

Entry of a Recombinant, Full-Length, Atoxic Tetanus Neurotoxin into Neuro-2a Cells

et al. 2014

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Tetanus neurotoxin (TeNT) and botulinum neurotoxin (BoNT) are clostridial neurotoxins (CNTs) that are the most toxic proteins for humans (1). TeNT and BoNT share ϳ35% identity and ϳ65% similarity and overall structure-function properties (2). BoNT intoxication results in flaccid paralysis through the inhibition of acetylcholine release by motor neurons, while TeNT intoxication yields a spastic paralysis due to inhibition of glycine release by inhibitory neurons (3). TeNT and BoNT are expressed as ϳ150-kDa single-chain proteins that are cleaved to form dichain proteins linked by a disulfide bond (2). The N-terminal 50-kDa light chain (LC) is a zinc-metalloprotease that cleaves neuronspecific soluble NSF attachment protein (SNAP) receptor (SNARE) proteins (4). TeNT and BoNT serotype B cleave the same residue within vesicle-associated membrane protein 2 (VAMP2), a SNARE protein of synaptic vesicles (SVs). The C-terminal 100-kDa heavy chain (HC) contains two structurally distinct domains with separate functions. The translocation domain (HCT) facilitates LC translocation from the SV lumen into the cell cytosol, and the receptor binding domain (HCR) binds dual host receptors.BoNT/A binds a ganglioside and synaptic vesicle protein 2 (SV2) and enters neurons upon SV recycling from the plasma membrane (5). Upon SV acidification within the periphery of motor neurons, the HCT undergoes a pH-dependent conformational change and inserts into the SV membrane, forming a channel that allows the LC to escape into the cytosol. Within the periphery of the motor neuron, the LC cleaves SNARE proteins, resulting in loss of stimulatory signaling between neurons and muscles, yielding flaccid paralysis. The LC of BoNT/A localizes to the plasma membrane to target synaptosomal-associated protein 25 (SNAP25) for cleavage within neurons (6).TeNT binds two gangliosides as functional receptors (7). TeNT can bind a glycophosphatidylinositol (GPI)-anchored protein (8) or SV2 (9), but the significance of these interactions has not been defined (10) or reproduced (11), respectively. TeNT enters motor neurons upon endocytosis (12) and traffics through motor neurons associated with Rab7-enriched endosomes that are of neutral pH (13,14). Retrograde trafficking proceeds from the axon to the soma, where TeNT transcytoses from the motor neuron into an inhibitory neuron of the central nervous system (CNS). Upon vesicle acidification, the LC is translocated into the cytosol and cleaves VAMP2. The block in signaling between the inhibitory neurons and motor neurons leads to the spastic paralysis characteristic of tetanus. The molecular mechanism responsible for the unique entry of BoNT and TeNT is not clearly understood.The modular structural domains of the CNTs have permitted the study of individual domains to assess protein structure-function in vitro; the LC is a functional SNARE protease (15), the HCT inserts into lipid bilayers and forms a channel (16), and the HCR binds host receptors (17). The tetanus HCR (HCR/T) can retrograde traffic in both cultu...

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Different response of the knockout mice lacking b-series gangliosides against botulinum and tetanus toxins

Cited by 38 publications

References 12 publications

Gangliosides as High Affinity Receptors for Tetanus Neurotoxin

Gangliosides as High Affinity Receptors for Tetanus Neurotoxin

Sialic acid-specific lectins: occurrence, specificity and function

Entry of a Recombinant, Full-Length, Atoxic Tetanus Neurotoxin into Neuro-2a Cells

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