Abstract
Using the yeast two-hybrid system with syntaxin-1A as bait, we isolated soluble NSF attachment protein (SNAP)-29 from a human brain cDNA library. Synaptosomal fractionation and immunocytochemical staining of hippocampal neurons in culture showed that SNAP-29 is present at synapses and is predominantly associated with synaptic vesicles. The interaction of SNAP-29 with syntaxin-1 was further confirmed with immunoprecipitation analysis. Binding competition studies with SNAP-29 demonstrated that it could compete with alpha-SNAP for binding to synaptic SNAP receptors (SNAREs) and consequently inhibit disassembly of the SNARE complex. Introduction of SNAP-29 into presynaptic superior cervical ganglion neurons in culture significantly inhibited synaptic transmission in an activity-dependent manner. Although SNAP-29 has been suggested to be a general SNARE component in membrane trafficking, our findings suggest that it may function as a regulator of SNARE complex disassembly and modulate the process of postfusion recycling of the SNARE components.
Publication types
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Research Support, Non-U.S. Gov't
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Research Support, U.S. Gov't, P.H.S.
MeSH terms
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Animals
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Antigens, Surface / genetics
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Antigens, Surface / metabolism
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Carrier Proteins / genetics
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Carrier Proteins / isolation & purification
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Carrier Proteins / metabolism
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Carrier Proteins / physiology*
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Cell Line, Transformed
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Cells, Cultured
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Hippocampus / cytology
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Hippocampus / metabolism
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Humans
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Membrane Proteins / metabolism*
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Nerve Tissue Proteins / genetics
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Nerve Tissue Proteins / metabolism*
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Neurons / metabolism
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Protein Binding
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Qb-SNARE Proteins
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Qc-SNARE Proteins
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Rats
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SNARE Proteins
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Soluble N-Ethylmaleimide-Sensitive Factor Attachment Proteins
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Subcellular Fractions
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Synaptic Transmission / physiology*
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Syntaxin 1
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Vesicular Transport Proteins*
Substances
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Antigens, Surface
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Carrier Proteins
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Membrane Proteins
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Nerve Tissue Proteins
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Qb-SNARE Proteins
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Qc-SNARE Proteins
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SNAP29 protein, human
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SNARE Proteins
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STX1A protein, human
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Soluble N-Ethylmaleimide-Sensitive Factor Attachment Proteins
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Stx1a protein, rat
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Syntaxin 1
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Vesicular Transport Proteins