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

• Published in: The Journal of immunology (1950) 2003-01, Vol.170 (2), p.1034-1042
• Main Authors: Mollinedo, Faustino, Martin-Martin, Belen, Calafat, Jero, Nabokina, Svetlana M, Lazo, Pedro A
• Format: Article
• Language: English
• Subjects: Antibodies, Monoclonal - pharmacology; Antigens, CD; Antigens, Neoplasm - metabolism; Arginine; Cell Adhesion Molecules - metabolism; Cell Membrane Permeability - immunology; Conserved Sequence; Cytoplasmic Granules - immunology; Cytoplasmic Granules - metabolism; Cytoplasmic Granules - ultrastructure; Electroporation; Exocytosis - immunology; Glutamine; GPI-Linked Proteins; Humans; Interphase - immunology; Membrane Proteins - classification; Membrane Proteins - immunology; Membrane Proteins - metabolism; Membrane Proteins - physiology; Membrane Proteins - ultrastructure; Microscopy, Immunoelectron; Molecular Sequence Data; Neutrophil Activation - immunology; Neutrophils - immunology; Neutrophils - metabolism; Neutrophils - ultrastructure; Qa-SNARE Proteins; R-SNARE Proteins; SNARE Proteins; Solubility; Tetanus Toxin - pharmacology; Vesicular Transport Proteins
• ISSN: 0022-1767; 1550-6606
• DOI: 10.4049/jimmunol.170.2.1034

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.