Involvement of SNAP-23 and syntaxin 6 in human neutrophil exocytosis.
To understand the molecular basis of exocytosis in human neutrophils, the role of syntaxin 6 and SNAP-23 in neutrophil degranulation was examined. Human syntaxin 6 was cloned and identified as a 255-amino acid protein with a carboxy-terminal transmembrane region and two coiled-coil domains. Syntaxin 6 was localized mainly in the plasma membrane of human resting neutrophils, whereas SNAP-23 was located primarily in the mobilizable tertiary and specific granules. SNAP-23 was translocated to the cell surface, colocalizing with syntaxin 6, on neutrophil activation. In vitro binding studies established that SNAP-23 binds to syntaxin 6. Coimmunoprecipitation assays indicated that SNAP-23 interacts with syntaxin 6 in vivo, and this interaction was dramatically increased on neutrophil activation. Antibodies against SNAP-23 inhibited Ca(++) and GTP-gamma-S-induced exocytosis of CD67-enriched specific granules, but they hardly affected exocytosis of the CD63-enriched azurophilic granules, when introduced into electropermeabilized neutrophils. Anti-syntaxin 6 antibodies prevented exocytosis of both CD67- and CD63-enriched granules in electropermeabilized neutrophils. These data show that syntaxin 6 and SNAP-23 are involved in human neutrophil exocytosis, demonstrating that vesicle SNAP receptor-target SNAP receptor (v-SNARE- t-SNARE) interactions modulate neutrophil secretion. Syntaxin 6 acts as a target for secretion of specific and azurophilic granules, whereas SNAP-23 mediates specific granule secretion.
Pubmed ID: 11001914 RIS Download
Amino Acid Sequence | Antibodies, Monoclonal | Antigens, CD | Antigens, CD63 | Antigens, Neoplasm | Calcium | Carrier Proteins | Cell Adhesion Molecules | Cell Degranulation | Cell Membrane | Cell Membrane Permeability | Cloning, Molecular | Exocytosis | Fluorescent Antibody Technique | Guanosine 5'-O-(3-Thiotriphosphate) | Humans | Immunoblotting | Membrane Glycoproteins | Membrane Proteins | Microscopy, Confocal | Molecular Sequence Data | Neutrophils | Platelet Membrane Glycoproteins | Qa-SNARE Proteins | Qb-SNARE Proteins | Qc-SNARE Proteins | RNA, Messenger | Reverse Transcriptase Polymerase Chain Reaction | Subcellular Fractions