PMID- 10413679
OWN - NLM
STAT- MEDLINE
DCOM- 19991028
LR  - 20220215
IS  - 0021-9533 (Print)
IS  - 0021-9533 (Linking)
VI  - 112 ( Pt 16)
DP  - 1999 Aug
TI  - Functional characterisation of tetanus and botulinum neurotoxins binding domains.
PG  - 2715-24
AB  - Tetanus and botulinum neurotoxins constitute a family of bacterial protein toxins
      responsible for two deadly syndromes in humans (tetanus and botulism,
      respectively). They bind with high affinity to neurons wherein they cause a
      complete inhibition of evoked neurotransmitter release. Here we report on the
      cloning, expression and use of the recombinant fragments of the heavy chains of
      tetanus neurotoxin and botulinum neurotoxin serotypes A, B and E as tools to
      study the neurospecific binding of the holotoxins. We found that the recombinant 
      50 kDa carboxy-terminal domains of tetanus and botulinum neurotoxins alone are
      responsible for the specific binding and internalisation into spinal cord cells
      in culture. Moreover, we provide evidence that the recombinant fragments block
      the internalization of the parental holotoxins in a dose-dependent manner, as
      determined by following the neurotoxin-dependent cleavage of their targets
      VAMP/synaptobrevin and SNAP-25. In addition, the recombinant binding fragments
      cause a significant delay in the paralysis induced by the corresponding holotoxin
      on the mouse phrenic nerve-hemidiaphragm preparation. Taken together, these
      results show that the carboxy-terminal domain of tetanus and botulinum
      neurotoxins is necessary and sufficient for the binding and internalisation of
      these proteins in neurons and open the possibility to use them as tools for the
      functional characterisation of the intracellular transport of clostridial
      neurotoxins.
FAU - Lalli, G
AU  - Lalli G
AD  - Molecular Neuropathobiology, Imperial Cancer Research Fund, London WC2A 3PX, UK.
FAU - Herreros, J
AU  - Herreros J
FAU - Osborne, S L
AU  - Osborne SL
FAU - Montecucco, C
AU  - Montecucco C
FAU - Rossetto, O
AU  - Rossetto O
FAU - Schiavo, G
AU  - Schiavo G
LA  - eng
SI  - GENBANK/AJ242628
GR  - 1068/TI_/Telethon/Italy
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
PL  - England
TA  - J Cell Sci
JT  - Journal of cell science
JID - 0052457
RN  - 0 (Membrane Proteins)
RN  - 0 (Nerve Tissue Proteins)
RN  - 0 (Peptide Fragments)
RN  - 0 (R-SNARE Proteins)
RN  - 0 (Recombinant Proteins)
RN  - 0 (SNARE Proteins)
RN  - 0 (Tetanus Toxin)
RN  - 0 (Vesicular Transport Proteins)
RN  - EC 3.4.24.69 (Botulinum Toxins)
SB  - IM
MH  - Animals
MH  - Binding Sites/genetics
MH  - Botulinum Toxins/*chemistry/genetics/*metabolism
MH  - Cells, Cultured
MH  - Dose-Response Relationship, Drug
MH  - Fetus/cytology
MH  - Gene Expression/physiology
MH  - Membrane Proteins/analysis/metabolism
MH  - Mice
MH  - Nerve Tissue Proteins/metabolism
MH  - Neuromuscular Junction/chemistry/metabolism
MH  - Neurons/chemistry/*metabolism
MH  - Peptide Fragments/genetics/metabolism
MH  - Phrenic Nerve/chemistry/cytology/metabolism
MH  - Protein Structure, Tertiary
MH  - R-SNARE Proteins
MH  - Recombinant Proteins/genetics/metabolism
MH  - SNARE Proteins
MH  - Spinal Cord/cytology
MH  - Tetanus Toxin/*chemistry/genetics/*metabolism
MH  - *Vesicular Transport Proteins
EDAT- 1999/07/22 00:00
MHDA- 1999/07/22 00:01
CRDT- 1999/07/22 00:00
PHST- 1999/07/22 00:00 [pubmed]
PHST- 1999/07/22 00:01 [medline]
PHST- 1999/07/22 00:00 [entrez]
AID - 10.1242/jcs.112.16.2715 [doi]
PST - ppublish
SO  - J Cell Sci. 1999 Aug;112 ( Pt 16):2715-24. doi: 10.1242/jcs.112.16.2715.