Synaptotagmin-7 phosphorylation mediates GLP-1-dependent potentiation of insulin secretion from β-cells

Proc Natl Acad Sci U S A. 2015 Aug 11;112(32):9996-10001. doi: 10.1073/pnas.1513004112. Epub 2015 Jul 27.

Abstract

Glucose stimulates insulin secretion from β-cells by increasing intracellular Ca(2+). Ca(2+) then binds to synaptotagmin-7 as a major Ca(2+) sensor for exocytosis, triggering secretory granule fusion and insulin secretion. In type-2 diabetes, insulin secretion is impaired; this impairment is ameliorated by glucagon-like peptide-1 (GLP-1) or by GLP-1 receptor agonists, which improve glucose homeostasis. However, the mechanism by which GLP-1 receptor agonists boost insulin secretion remains unclear. Here, we report that GLP-1 stimulates protein kinase A (PKA)-dependent phosphorylation of synaptotagmin-7 at serine-103, which enhances glucose- and Ca(2+)-stimulated insulin secretion and accounts for the improvement of glucose homeostasis by GLP-1. A phospho-mimetic synaptotagmin-7 mutant enhances Ca(2+)-triggered exocytosis, whereas a phospho-inactive synaptotagmin-7 mutant disrupts GLP-1 potentiation of insulin secretion. Our findings thus suggest that synaptotagmin-7 is directly activated by GLP-1 signaling and may serve as a drug target for boosting insulin secretion. Moreover, our data reveal, to our knowledge, the first physiological modulation of Ca(2+)-triggered exocytosis by direct phosphorylation of a synaptotagmin.

Keywords: diabetes; exocytosis; incretin; phosphorylation; synaptotagmin.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Sequence
  • Animals
  • Colforsin / pharmacology
  • Conserved Sequence
  • Cyclic AMP / pharmacology
  • Cyclic AMP-Dependent Protein Kinases / metabolism
  • Evolution, Molecular
  • Exenatide
  • Exocytosis / drug effects
  • Glucagon-Like Peptide 1 / metabolism*
  • Glucagon-Like Peptide-1 Receptor
  • Glucose / pharmacology
  • HEK293 Cells
  • Humans
  • Insulin / metabolism*
  • Insulin Secretion
  • Insulin-Secreting Cells / drug effects
  • Insulin-Secreting Cells / metabolism*
  • Mice, Knockout
  • Molecular Sequence Data
  • Mutation / genetics
  • Peptides / pharmacology
  • Phosphorylation / drug effects
  • Phosphoserine / metabolism
  • Rats
  • Receptors, Glucagon / metabolism
  • Synaptotagmins / chemistry
  • Synaptotagmins / metabolism*
  • Venoms / pharmacology

Substances

  • GLP1R protein, human
  • Glp1r protein, mouse
  • Glp1r protein, rat
  • Glucagon-Like Peptide-1 Receptor
  • Insulin
  • Peptides
  • Receptors, Glucagon
  • Venoms
  • Synaptotagmins
  • Phosphoserine
  • Colforsin
  • Glucagon-Like Peptide 1
  • Exenatide
  • Cyclic AMP
  • Cyclic AMP-Dependent Protein Kinases
  • Glucose