Vesicle associated membrane protein (VAMP)-7 and VAMP-8, but not VAMP-2 or VAMP-3, are required for activation-induced degranulation of mature human mast cells.
Mediator release from mast cells (MC) is a crucial step in allergic and non-allergic inflammatory disorders. However, the final events in response to activation leading to membrane fusion and thereby facilitating degranulation have hitherto not been analyzed in human MC. Soluble N-ethyl-maleimide-sensitive factor attachment protein receptors (SNARE) represent a highly conserved family of proteins that have been shown to mediate intracellular membrane fusion events. Here, we show that mature MC isolated from human intestinal tissue express soluble N-ethylmaleide sensitive factor attachment protein (SNAP)-23, Syntaxin (STX)-1B, STX-2, STX-3, STX-4, and STX-6 but not SNAP-25. Furthermore, we found that primary human MC express substantial amounts of vesicle associated membrane protein (VAMP)-3, VAMP-7 and VAMP-8 and, in contrast to previous reports about rodent MC, only low levels of VAMP-2. Furthermore, VAMP-7 and VAMP-8 were found to translocate to the plasma membrane and interact with SNAP-23 and STX-4 upon activation. Inhibition of SNAP-23, STX-4, VAMP-7 or VAMP-8, but not VAMP-2 or VAMP-3, resulted in a markedly reduced high-affinity IgE receptor-mediated histamine release. In summary, our data show that mature human MC express a specific pattern of SNARE and that VAMP-7 and VAMP-8, but not VAMP-2, are required for rapid degranulation.
Sander
Leif E
LE
Institute of Nutritional Medicine, University of Hohenheim, Stuttgart, Germany.
Frank
Simon P C
SP
Bolat
Seza
S
Blank
Ulrich
U
Galli
Thierry
T
Bigalke
Hans
H
Bischoff
Stephan C
SC
Lorentz
Axel
A
D000906:Antibodies / Q000494:pharmacology; D015153:Blotting, Western; D015550:Cell Degranulation / Q000187:drug effects / Q000502:physiology*; D002462:Cell Membrane / Q000378:metabolism; D003593:Cytoplasm / Q000378:metabolism; D005455:Fluorescent Antibody Technique; D015870:Gene Expression / Q000187:drug effects; D006636:Histamine Release / Q000187:drug effects / Q000502:physiology; D006801:Humans; D008407:Mast Cells / Q000187:drug effects / Q000378:metabolism / Q000502:physiology*; D008666:Metalloendopeptidases / Q000494:pharmacology; D050765:Qa-SNARE Proteins / Q000235:genetics / Q000276:immunology / Q000378:metabolism; D050766:Qb-SNARE Proteins / Q000235:genetics / Q000276:immunology / Q000378:metabolism; D050767:Qc-SNARE Proteins / Q000235:genetics / Q000276:immunology / Q000378:metabolism; D050683:R-SNARE Proteins / Q000235:genetics / Q000378:metabolism / Q000502:physiology*; D020133:Reverse Transcriptase Polymerase Chain Reaction; D050600:SNARE Proteins / Q000235:genetics / Q000378:metabolism; D013744:Tetanus Toxin / Q000494:pharmacology; D050988:Vesicle-Associated Membrane Protein 2 / Q000235:genetics / Q000378:metabolism / Q000502:physiology; D051040:Vesicle-Associated Membrane Protein 3 / Q000235:genetics / Q000378:metabolism / Q000502:physiology
D000906:Antibodies; D050765:Qa-SNARE Proteins; D050766:Qb-SNARE Proteins; D050767:Qc-SNARE Proteins; D050683:R-SNARE Proteins; C100493:SNAP23 protein, human; D050600:SNARE Proteins; D013744:Tetanus Toxin; C494807:VAMP2 protein, human; C494946:VAMP3 protein, human; C121241:VAMP7 protein, human; C112938:VAMP8 protein, human; D050988:Vesicle-Associated Membrane Protein 2; D051040:Vesicle-Associated Membrane Protein 3; D008666:Metalloendopeptidases; C093556:zinc-endopeptidase, tetanus neurotoxin