Tetanus toxin receptor Specific cross‐linking of tetanus toxin to a protein of NGF‐differentiated PC 12 cells

Schiavo, Giampietro; Ferrari, Giovanna; Rossetto, Ornella; Montecucco, Cesare

doi:10.1016/0014-5793(91)81266-b

Cited by 43 publications

(19 citation statements)

References 43 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Furthermore, chemical treatment with periodic acid has been used to enhance the labeling of sialoglycocomponents by radiodinated tetanus toxin, a related clostridial neurotoxin, in NGF-differentiated rat pheochromocytoma cell line (Nathan and Yavin, 1989). Moreover, using chemical cross-linking agents and NGF-differentiated sub-clone of these cells it has been described the interaction of tetanus toxin with a receptor protein (Schiavo et al, 1991).…”

Section: Introductionmentioning

confidence: 98%

Binding of botulinum neurotoxin to pure cholinergic nerve terminals isolated from the electric organ of Torpedo

Blasi

Egea

Castiella

et al. 1992

J. Neural Transmission

View full text Add to dashboard Cite

Torpedo electric organ has been used to study the binding of botulinum neurotoxin type A to pure cholinergic synaptosomes and presynaptic plasma membrane. 125I-labeled botulinum neurotoxin type A exhibits specific binding to cholinergic fractions. Two binding sites have been determined according to data analysis: a high affinity binding site (synaptosomes: Kd = 0.11 +/- 0.03 nM, Bmax = 50 +/- 10 fmol.mg prot-1; presynaptic plasma membrane: Kd = 0.2 +/- 0.05 nM, Bmax = 150 +/- 15 fmol.mg prot-1) and a low affinity binding site (synaptosomes: Kd approximately 26 nM, Bmax approximately 7.5 pmol.mg prot-1; presynaptic plasma membrane: Kd approximately 30 nM, Bmax approximately 52 pmol.mg prot-1). The binding of 125I-botulinum neurotoxin type A is decreased by previous treatment of synaptosomes by neuraminidase and trypsin, and by a preincubation with bovine brain gangliosides or antiserum raised against Torpedo presynaptic plasma membrane. When presynaptic plasma membranes are blotted to nitrocellulose sheet, either 125I-botulinum neurotoxin or botulinum toxin-gold complexes bind to a M(r) approximately 140,000 protein. Botulinum toxin-gold complexes have also been used to study the toxin internalization process into Torpedo synaptosomes. The images fit the three step sequence model in the pathway of botulinum neurotoxin poisoning.

show abstract

Section: Introductionmentioning

confidence: 98%

Binding of botulinum neurotoxin to pure cholinergic nerve terminals isolated from the electric organ of Torpedo

Blasi

Egea

Castiella

et al. 1992

J. Neural Transmission

View full text Add to dashboard Cite

show abstract

“…TeNT was suggested to interact with the luminal domain of a synaptic vesicle protein in central synapses (Matteoli et al, 1996). Crosslinking experiments performed on nerve growth factor (NGF)‐differentiated PC12 cells with native TeNT suggested the presence of a putative 20‐kDa receptor protein for this neurotoxin (Schiavo et al, 1991). However, no further characterisation of this protein has yet been provided.…”

mentioning

confidence: 99%

Tetanus Toxin Fragment C Binds to a Protein Present in Neuronal Cell Lines and Motoneurons

Herreros¹,

Lalli²,

Montecucco³

et al. 2000

Journal of Neurochemistry

Self Cite

View full text Add to dashboard Cite

Abstract:Tetanus neurotoxin is one of the most powerful protein toxins known, acting in vivo at femtomolar doses. Two main factors determine its high potency: a protease activity restricted to a single intracellular substrate and its absolute neurospecificity. Whereas the enzymatic properties of tetanus toxin have been thoroughly defined, the nature of its neuronal receptor(s) and their involvement in the intracellular trafficking of tetanus toxin are poorly understood. Using binding and crosslinking experiments, we report here on the characterisation of an N-glycosylated 15-kDa interacting protein, which behaves as an integral membrane protein. This putative receptor specifically interacts with the binding domain (fragment C) of tetanus toxin and not with several related botulinum neurotoxins in spinal cord motoneurons and neuronal-like cell lines. Sialic acid-specific lectins antagonise the binding of tetanus toxin to the cell surface and to the 15-kDa protein, supporting the central role of sialic acid residues in the recognition process. Altogether, these results indicate the existence of a neuronal protein receptor for tetanus toxin whose identification is likely to constitute a key step in the analysis of the molecular machinery involved in the toxin internalisation and retrograde transport.

show abstract

“…Several BoNT serotypes interact with synaptotagmins (Nishiki et al, 1994;Li and Singh, 1998), but the role of these proteins as physiological BoNT receptors remains controversial (Evans et al, 1986;Bakry et al, 1997). We have previously followed a cross-linking approach (Schiavo et al, 1991) to demonstrate that the binding domain of TeNT (TeNT H C ) interacts with a glycoprotein of ϳ15 kDa (p15) in nerve growth factor (NGF)-differentiated PC12 cells and spinal cord MNs (Her-reros et al, 2000a). In these neuronal cell types p15 showed the behavior of an integral membrane protein (Herreros et al, 2000a).…”

Section: Introductionmentioning

confidence: 99%

Lipid Rafts Act as Specialized Domains for Tetanus Toxin Binding and Internalization into Neurons

Herreros

Schiavo

2001

MBoC

Self Cite

161

122

View full text Add to dashboard Cite

Tetanus (TeNT) is a zinc protease that blocks neurotransmission by cleaving the synaptic protein vesicle-associated membrane protein/synaptobrevin. Although its intracellular catalytic activity is well established, the mechanism by which this neurotoxin interacts with the neuronal surface is not known. In this study, we characterize p15s, the first plasma membrane TeNT binding proteins and we show that they are glycosylphosphatidylinositol-anchored glycoproteins in nerve growth factor (NGF)-differentiated PC12 cells, spinal cord cells, and purified motor neurons. We identify p15 as neuronal Thy-1 in NGF-differentiated PC12 cells. Fluorescence lifetime imaging microscopy measurements confirm the close association of the binding domain of TeNT and Thy-1 at the plasma membrane. We find that TeNT is recruited to detergent-insoluble lipid microdomains on the surface of neuronal cells. Finally, we show that cholesterol depletion affects a raft subpool and blocks the internalization and intracellular activity of the toxin. Our results indicate that TeNT interacts with target cells by binding to lipid rafts and that cholesterol is required for TeNT internalization and/or trafficking in neurons. INTRODUCTIONTetanus (TeNT) and botulinum neurotoxins (BoNTs) block neurotransmitter release and are responsible for tetanus and botulism, respectively. These toxins share a common structure comprised of a heavy (H, 100 kDa) and a light (L, 50 kDa) chain linked by a disulfide bond. The H chain mediates binding and internalization in neurons, whereas the L chain is a metallo-protease that selectively cleaves synaptic proteins (Niemann et al., 1994;Schiavo et al., 2000). TeNT and BoNTs bind to the neuromuscular junction, but their intracellular actions take place at different levels of the nervous system. TeNT undergoes retrograde transport to the cell body of spinal cord motor neurons (MNs), is transcytosed, and cleaves the synaptic vesicle protein vesicle-associated membrane protein (VAMP)/synaptobrevin in inhibitory synapses. Instead, BoNTs act at a peripheral level by blocking acetylcholine release at the motor nerve terminal. This differential sorting has been interpreted as a consequence of binding to different surface receptors (Habermann and Dreyer, 1986;Herreros et al., 1999).TeNT and BoNTs bind to polysialogangliosides of the G1b series (Halpern and Neale, 1995). However, the fact that their binding is sensitive to proteases (Lazarovici and Yavin, 1986;Pierce et al., 1986;Yavin and Nathan, 1986) suggests the existence of specific protein receptors. Thus, a model in which TeNT and BoNTs interact with a complex constituted by both lipid and protein receptors has been proposed (Montecucco, 1986). Despite several efforts, these protein receptors have not been conclusively identified. Several BoNT serotypes interact with synaptotagmins (Nishiki et al., 1994;Li and Singh, 1998), but the role of these proteins as physiological BoNT receptors remains controversial (Evans et al., 1986;Bakry et al., 1997). We have previously followe...

show abstract

Tetanus toxin receptor Specific cross‐linking of tetanus toxin to a protein of NGF‐differentiated PC 12 cells

Cited by 43 publications

References 43 publications

Binding of botulinum neurotoxin to pure cholinergic nerve terminals isolated from the electric organ of Torpedo

Binding of botulinum neurotoxin to pure cholinergic nerve terminals isolated from the electric organ of Torpedo

Tetanus Toxin Fragment C Binds to a Protein Present in Neuronal Cell Lines and Motoneurons

Lipid Rafts Act as Specialized Domains for Tetanus Toxin Binding and Internalization into Neurons

Contact Info

Product

Resources

About