Tetanus Toxin Inhibits Depolarization‐Stimulated Protein Phosphorylation in Rat Cortical Synaptosomes: Effect on Synapsin I Phosphorylation and Translocation

Peter Presek, Sönke Jessen, Florian Dreyer, Paula E. Jarvie, Duygu Findik, Peter R. Dunkley

Research output: Contribution to journalArticle

23 Citations (Scopus)

Abstract

Abstract: Synapsin I, a prominent phosphoprotein in nerve terminals, is proposed to modulate exocytosis by interaction with the cytoplasmic surface of small synaptic vesicles and cytoskeletal elements in a phosphorylation‐dependent manner. Tetanus toxin (TeTx), a potent inhibitor of neurotransmitter release, attenuated the depolarization‐stimulated increase in synapsin I phosphorylation in rat cortical particles and in synaptosomes. TeTx also markedly decreased the translocation of synapsin I from the small synaptic vesicles and the cytoskeleton into the cytosol, on depolarization of synaptosomes. The effect of TeTx on synapsin I phosphorylation was both time and TeTx concentration dependent and required active toxin. One‐ and two‐dimensional peptide maps of synapsin I with V8 proteinase and trypsin, respectively, showed no differences in the relative phosphorylation of peptides for the control and TeTx‐treated synaptosomes, suggesting that both the calmodulin‐and the cyclic AMP‐dependent kinases that label this protein are equally affected. Phosphorylation of synapsin IIb and the B‐50 protein (GAP43), a known substrate of protein kinase C, was also inhibited by TeTx. TeTx affected only a limited number of phosphoproteins and the calcium‐dependent decrease in dephosphin phosphorylation remained unaffected. In vitro phosphorylation of proteins in lysed synaptosomes was not influenced by prior TeTx treatment of the intact synaptosomes or by the addition of TeTx to lysates, suggesting that the effect of TeTx on protein phosphorylation was indirect. Our data demonstrate that TeTx inhibits neurotransmitter release, the phosphorylation of a select group of phosphoproteins in nerve terminals, and the translocation of synapsin I. These findings contribute to our understanding of the basic mechanism of TeTx action.

Original languageEnglish (US)
Pages (from-to)1336-1343
Number of pages8
JournalJournal of Neurochemistry
Volume59
Issue number4
DOIs
StatePublished - Oct 1992

Fingerprint

Synapsins
Tetanus Toxin
Phosphorylation
Synaptosomes
Rats
Proteins
Phosphoproteins
Synaptic Vesicles
Neurotransmitter Agents
Dynamins
GAP-43 Protein
Peptides
Exocytosis
Depolarization
Cytoskeleton
Cytosol
Trypsin
Protein Kinases
Protein Kinase C
Labels

Keywords

  • Depolarization
  • Synapsin I phosphorylation
  • Synapsin I translocation
  • Synaptosomes (rat)
  • Tetanus toxin

ASJC Scopus subject areas

  • Biochemistry
  • Cellular and Molecular Neuroscience

Cite this

Tetanus Toxin Inhibits Depolarization‐Stimulated Protein Phosphorylation in Rat Cortical Synaptosomes : Effect on Synapsin I Phosphorylation and Translocation. / Presek, Peter; Jessen, Sönke; Dreyer, Florian; Jarvie, Paula E.; Findik, Duygu; Dunkley, Peter R.

In: Journal of Neurochemistry, Vol. 59, No. 4, 10.1992, p. 1336-1343.

Research output: Contribution to journalArticle

Presek, Peter ; Jessen, Sönke ; Dreyer, Florian ; Jarvie, Paula E. ; Findik, Duygu ; Dunkley, Peter R. / Tetanus Toxin Inhibits Depolarization‐Stimulated Protein Phosphorylation in Rat Cortical Synaptosomes : Effect on Synapsin I Phosphorylation and Translocation. In: Journal of Neurochemistry. 1992 ; Vol. 59, No. 4. pp. 1336-1343.
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abstract = "Abstract: Synapsin I, a prominent phosphoprotein in nerve terminals, is proposed to modulate exocytosis by interaction with the cytoplasmic surface of small synaptic vesicles and cytoskeletal elements in a phosphorylation‐dependent manner. Tetanus toxin (TeTx), a potent inhibitor of neurotransmitter release, attenuated the depolarization‐stimulated increase in synapsin I phosphorylation in rat cortical particles and in synaptosomes. TeTx also markedly decreased the translocation of synapsin I from the small synaptic vesicles and the cytoskeleton into the cytosol, on depolarization of synaptosomes. The effect of TeTx on synapsin I phosphorylation was both time and TeTx concentration dependent and required active toxin. One‐ and two‐dimensional peptide maps of synapsin I with V8 proteinase and trypsin, respectively, showed no differences in the relative phosphorylation of peptides for the control and TeTx‐treated synaptosomes, suggesting that both the calmodulin‐and the cyclic AMP‐dependent kinases that label this protein are equally affected. Phosphorylation of synapsin IIb and the B‐50 protein (GAP43), a known substrate of protein kinase C, was also inhibited by TeTx. TeTx affected only a limited number of phosphoproteins and the calcium‐dependent decrease in dephosphin phosphorylation remained unaffected. In vitro phosphorylation of proteins in lysed synaptosomes was not influenced by prior TeTx treatment of the intact synaptosomes or by the addition of TeTx to lysates, suggesting that the effect of TeTx on protein phosphorylation was indirect. Our data demonstrate that TeTx inhibits neurotransmitter release, the phosphorylation of a select group of phosphoproteins in nerve terminals, and the translocation of synapsin I. These findings contribute to our understanding of the basic mechanism of TeTx action.",
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AU - Presek, Peter

AU - Jessen, Sönke

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AU - Jarvie, Paula E.

AU - Findik, Duygu

AU - Dunkley, Peter R.

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N2 - Abstract: Synapsin I, a prominent phosphoprotein in nerve terminals, is proposed to modulate exocytosis by interaction with the cytoplasmic surface of small synaptic vesicles and cytoskeletal elements in a phosphorylation‐dependent manner. Tetanus toxin (TeTx), a potent inhibitor of neurotransmitter release, attenuated the depolarization‐stimulated increase in synapsin I phosphorylation in rat cortical particles and in synaptosomes. TeTx also markedly decreased the translocation of synapsin I from the small synaptic vesicles and the cytoskeleton into the cytosol, on depolarization of synaptosomes. The effect of TeTx on synapsin I phosphorylation was both time and TeTx concentration dependent and required active toxin. One‐ and two‐dimensional peptide maps of synapsin I with V8 proteinase and trypsin, respectively, showed no differences in the relative phosphorylation of peptides for the control and TeTx‐treated synaptosomes, suggesting that both the calmodulin‐and the cyclic AMP‐dependent kinases that label this protein are equally affected. Phosphorylation of synapsin IIb and the B‐50 protein (GAP43), a known substrate of protein kinase C, was also inhibited by TeTx. TeTx affected only a limited number of phosphoproteins and the calcium‐dependent decrease in dephosphin phosphorylation remained unaffected. In vitro phosphorylation of proteins in lysed synaptosomes was not influenced by prior TeTx treatment of the intact synaptosomes or by the addition of TeTx to lysates, suggesting that the effect of TeTx on protein phosphorylation was indirect. Our data demonstrate that TeTx inhibits neurotransmitter release, the phosphorylation of a select group of phosphoproteins in nerve terminals, and the translocation of synapsin I. These findings contribute to our understanding of the basic mechanism of TeTx action.

AB - Abstract: Synapsin I, a prominent phosphoprotein in nerve terminals, is proposed to modulate exocytosis by interaction with the cytoplasmic surface of small synaptic vesicles and cytoskeletal elements in a phosphorylation‐dependent manner. Tetanus toxin (TeTx), a potent inhibitor of neurotransmitter release, attenuated the depolarization‐stimulated increase in synapsin I phosphorylation in rat cortical particles and in synaptosomes. TeTx also markedly decreased the translocation of synapsin I from the small synaptic vesicles and the cytoskeleton into the cytosol, on depolarization of synaptosomes. The effect of TeTx on synapsin I phosphorylation was both time and TeTx concentration dependent and required active toxin. One‐ and two‐dimensional peptide maps of synapsin I with V8 proteinase and trypsin, respectively, showed no differences in the relative phosphorylation of peptides for the control and TeTx‐treated synaptosomes, suggesting that both the calmodulin‐and the cyclic AMP‐dependent kinases that label this protein are equally affected. Phosphorylation of synapsin IIb and the B‐50 protein (GAP43), a known substrate of protein kinase C, was also inhibited by TeTx. TeTx affected only a limited number of phosphoproteins and the calcium‐dependent decrease in dephosphin phosphorylation remained unaffected. In vitro phosphorylation of proteins in lysed synaptosomes was not influenced by prior TeTx treatment of the intact synaptosomes or by the addition of TeTx to lysates, suggesting that the effect of TeTx on protein phosphorylation was indirect. Our data demonstrate that TeTx inhibits neurotransmitter release, the phosphorylation of a select group of phosphoproteins in nerve terminals, and the translocation of synapsin I. These findings contribute to our understanding of the basic mechanism of TeTx action.

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