Association of syntaxin with SNAP 25 and VAMP (synaptobrevin) in Torpedo synaptosomes.
Two proteins of the presynaptic plasma membrane, syntaxin and SNAP 25, and VAMP/ synaptobrevin, a synaptic vesicle membrane protein, form stable protein complexes which are involved in the docking and fusion of synaptic vesicles at the mammalian brain presynaptic membrane. Similar protein complexes were revealed in an homogeneous population of cholinergic synaptosomes purified from Torpedo electric organ by combining velocity sedimentation and immunoprecipitation experiments. After CHAPS solubilization, virtually all the nerve terminal syntaxin was found in the form of large 16 S complexes, in association with 65% of SNAP 25 and 15% of VAMP. Upon Triton X100 solubilization, syntaxin was still recovered in association with SNAP 25 and VAMP but in smaller 8 S complexes. A small (2-5%) percentage of the nerve terminal 15 kDa proteolipid subunit of the v-H+ATPase and of mediatophore was copurified with syntaxin, using two different antisyntaxin monoclonal antibodies. The use of an homogeneous population of peripheral cholinergic nerve terminals allowed us to extend results on the composition of the brain presynaptic protein complexes to the Torpedo electric organ synapse, a model of the rapid neuromuscular synapses.
Shiff
G
G
Laboratoire de Neurobiologie Cellulaire et Moleculaire, C.N.R.S., 91198, Gif sur Yvette, France.
Synguelakis
M
M
Morel
N
N
D000109:Acetylcholine / Q000502:physiology*; D000818:Animals; D000911:Antibodies, Monoclonal; D001923:Brain Chemistry / Q000502:physiology; D004557:Electric Organ / Q000737:chemistry*; D008565:Membrane Proteins / Q000737:chemistry; D008959:Models, Neurological; D009046:Motor Neurons / Q000737:chemistry; D009411:Nerve Endings / Q000737:chemistry*; D009419:Nerve Tissue Proteins / Q000737:chemistry*; D011233:Precipitin Tests; D050765:Qa-SNARE Proteins; D050683:R-SNARE Proteins; D050825:Synaptosomal-Associated Protein 25; D013574:Synaptosomes / Q000737:chemistry*; D014101:Torpedo
D000911:Antibodies, Monoclonal; D008565:Membrane Proteins; D009419:Nerve Tissue Proteins; D050765:Qa-SNARE Proteins; D050683:R-SNARE Proteins; D050825:Synaptosomal-Associated Protein 25; D000109:Acetylcholine