Role of vesicle-associated membrane protein-2, through Q-soluble N-ethylmaleimide-sensitive factor attachment protein receptor/R-soluble N-ethylmaleimide-sensitive factor attachment protein receptor interaction, in the exocytosis of specific and tertiary granules of human neutrophils.
We have examined the role of the R-soluble N-ethylmaleimide-sensitive factor attachment protein receptor (SNARE) synaptobrevin-2/vesicle-associated membrane protein (VAMP)-2 in neutrophil exocytosis. VAMP-2, localized in the membranes of specific and gelatinase-containing tertiary granules in resting human neutrophils, resulted translocated to the cell surface following neutrophil activation under experimental conditions that induced ... exocytosis of specific and tertiary granules. VAMP-2 was also found on the external membrane region of granules docking to the plasma membrane in activated neutrophils. Specific Abs against VAMP-2 inhibited Ca(2+) and GTP-gamma-S-induced exocytosis of CD66b-enriched specific and tertiary granules, but did not affect exocytosis of CD63-enriched azurophilic granules, in electropermeabilized neutrophils. Tetanus toxin disrupted VAMP-2 and inhibited exocytosis of tertiary and specific granules. Activation of neutrophils led to the interaction of VAMP-2 with the plasma membrane Q-SNARE syntaxin 4, and anti-syntaxin 4 Abs inhibited exocytosis of specific and tertiary granules in electropermeabilized neutrophils. Immunoelectron microscopy showed syntaxin 4 on the plasma membrane contacting with docked granules in activated neutrophils. These data indicate that VAMP-2 mediates exocytosis of specific and tertiary granules, and that Q-SNARE/R-SNARE complexes containing VAMP-2 and syntaxin 4 are involved in neutrophil exocytosis.
Mesh Terms:
Antibodies, Monoclonal, Antigens, CD, Antigens, Neoplasm, Arginine, Cell Adhesion Molecules, Cell Membrane Permeability, Conserved Sequence, Cytoplasmic Granules, Electroporation, Exocytosis, Glutamine, Humans, Interphase, Membrane Proteins, Microscopy, Immunoelectron, Molecular Sequence Data, Neutrophil Activation, Neutrophils, Qa-SNARE Proteins, R-SNARE Proteins, SNARE Proteins, Solubility, Tetanus Toxin, Vesicular Transport Proteins
Antibodies, Monoclonal, Antigens, CD, Antigens, Neoplasm, Arginine, Cell Adhesion Molecules, Cell Membrane Permeability, Conserved Sequence, Cytoplasmic Granules, Electroporation, Exocytosis, Glutamine, Humans, Interphase, Membrane Proteins, Microscopy, Immunoelectron, Molecular Sequence Data, Neutrophil Activation, Neutrophils, Qa-SNARE Proteins, R-SNARE Proteins, SNARE Proteins, Solubility, Tetanus Toxin, Vesicular Transport Proteins
J. Immunol.
Date: Jan. 15, 2003
PubMed ID: 12517971
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