Stimulus-induced S-nitrosylation of syntaxin 4 impacts insulin granule exocytosis

Dean A. Wiseman, Michael A. Kalwat, Debbie C. Thurmond

Research output: Contribution to journalArticle

32 Citations (Scopus)

Abstract

Glucose-stimulated insulin release from pancreatic islet β-cells involves increased levels of reactive oxygen and nitrogen species. Although this is normal, under pathophysiological conditions such as chronic hyperglycemia and inflammation, insulin exocytosis fails, and yet the mechanistic reason for failure is unclear. Hypothesizing that exocytotic proteins might be targets of S-nitrosylation, with their dysfunction under conditions of nitrosative stress serving as a mechanistic basis for insulin secretory dysfunction, we identified the t-SNARE protein Syntaxin 4 as a target of modification by S-nitrosylation. The cellular content of S-nitrosylated Syntaxin 4 peaked acutely, within 5 min of glucose stimulation in both human islets and MIN6 β-cells, corresponding to the time at which Syntaxin 4 activation was detectable. S-Nitrosylation was mapped to Syntaxin 4 residue Cys141, located within the Hc domain predicted to increase accessibility for v-SNARE interaction. A C141S-Syntaxin 4 mutant resisted S-nitrosylation induced in vitro by the nitric oxide donor compound S-nitroso-L-glutathione, failed to exhibit glucose-induced activation and VAMP2 binding, and failed to potentiate insulin release akin to that of wild-type Syntaxin 4. Strikingly, S-nitrosylation of Syntaxin 4 could be induced by acute treatment with inflammatory cytokines (TNFα, IL-1β, and IFNγ), coordinate with inappropriate Syntaxin 4 activation and insulin release in the absence of the glucose stimulus, consistent with nitrosative stress and dysfunctional exocytosis, preceding the cell dysfunction and death associated with more chronic stimulation (24 h). Taken together, these data indicate a significant role for reactive nitrogen species in the insulin exocytosis mechanism in β-cells and expose a potential pathophysiological exploitation of this mechanism to underlie dysfunctional exocytosis.

Original languageEnglish
Pages (from-to)16344-16354
Number of pages11
JournalJournal of Biological Chemistry
Volume286
Issue number18
DOIs
StatePublished - May 6 2011

Fingerprint

Qa-SNARE Proteins
Exocytosis
Insulin
SNARE Proteins
Islets of Langerhans
Reactive Nitrogen Species
Glucose
Chemical activation
Vesicle-Associated Membrane Protein 2
Nitroso Compounds
Nitric Oxide Donors
Interleukin-1
Hyperglycemia
Glutathione
Reactive Oxygen Species
Cell Death
Cytokines
Inflammation

ASJC Scopus subject areas

  • Biochemistry
  • Cell Biology
  • Molecular Biology

Cite this

Stimulus-induced S-nitrosylation of syntaxin 4 impacts insulin granule exocytosis. / Wiseman, Dean A.; Kalwat, Michael A.; Thurmond, Debbie C.

In: Journal of Biological Chemistry, Vol. 286, No. 18, 06.05.2011, p. 16344-16354.

Research output: Contribution to journalArticle

Wiseman, Dean A. ; Kalwat, Michael A. ; Thurmond, Debbie C. / Stimulus-induced S-nitrosylation of syntaxin 4 impacts insulin granule exocytosis. In: Journal of Biological Chemistry. 2011 ; Vol. 286, No. 18. pp. 16344-16354.
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