@article{b9a729f0fb7111de825d000ea68e967b,
title = "Protein kinase C-dependent phosphorylation of synaptosome-associated protein of 25 kDa at Ser187 potentiates vesicle recruitment",
abstract = "Activation of protein kinase C (PKC) constitutes a key event in the upregulation of secretory strength in neurons and neurosecretory cells during extensive stimulation, presumably by speeding up vesicle supply. However, the molecular targets and their mode of action remain elusive. We studied the only PKC-dependent phosphorylation site in the neuronal soluble N-ethylmaleimide-sensitive factor attachment protein receptor (SNARE) complex, Ser(187), in synaptosome-associated protein of 25 kDa (SNAP-25). This phosphorylation site is located within the negatively charged C-terminal end of SNAP-25, which has been shown to be of critical importance in calcium-triggered exocytosis. We combined mutational studies that used overexpression in chromaffin cells with capacitance measurements and flash photolysis of caged calcium, allowing for high time resolution during both the stimulation and measurement of exocytosis. Overexpression of mutants simulating the phosphorylated form of Ser(187) accelerated vesicle recruitment after the emptying of the releasable vesicle pools. Overexpression of mutants simulating the nonphosphorylated form, or block of PKC, impaired the refilling of the vesicle pools to similar extents. Biochemical studies verified the phosphorylation of a subpopulation of SNAP-25 after elevation of intracellular calcium concentrations. Some of the mutations led to a moderately decreased fast exocytotic burst component, which did not seem to be associated with the phosphorylation state of SNAP-25. Thus the C terminus of SNAP-25 plays a role for both fast exocytosis triggering and vesicle recruitment, and the latter process is regulated by PKC-dependent phosphorylation.",
author = "G{\'a}bor Nagy and Ulf Matti and Nehring, {Ralf B} and Thomas Binz and Jens Rettig and Erwin Neher and S{\o}rensen, {Jakob B}",
note = "Keywords: Animals; Antibodies; Calcium; Cattle; Cell Membrane; Cells, Cultured; Chromaffin Cells; Computer Simulation; Electric Capacitance; Enzyme Inhibitors; Exocytosis; Fluorescent Dyes; Green Fluorescent Proteins; Luminescent Proteins; Membrane Proteins; Mutagenesis, Site-Directed; Nerve Tissue Proteins; Patch-Clamp Techniques; Phosphorylation; Photolysis; Protein Kinase C; Recombinant Fusion Proteins; SNARE Proteins; Secretory Vesicles; Structure-Activity Relationship; Synaptosomal-Associated Protein 25; Vesicular Transport Proteins",
year = "2002",
language = "English",
volume = "22",
pages = "9278--86",
journal = "Journal of Neuroscience",
issn = "0270-6474",
publisher = "Society for Neuroscience",
number = "21",
}