Physical link and functional coupling of presynaptic calcium channels and the synaptic vesicle docking/fusion machinery.
N- and P/Q-type calcium channels are localized in high density in presynaptic nerve terminals and are crucial elements in neuronal excitation-secretion coupling. In addition to mediating Ca2+ entry to initiate transmitter release, they are thought to interact directly with proteins of the synaptic vesicle docking/fusion machinery. As outlined in the preceding article, these calcium channels can be purified from brain as a complex with SNARE proteins which are involved in exocytosis. In addition, N-type and P/Q-type calcium channels are co-localized with syntaxin in high-density clusters in nerve terminals. Here we review the role of the synaptic protein interaction (synprint) sites in the intracellular loop II-III (L(II-III)) of both alpha1B and alpha1A subunits of N-type and P/Q-type calcium channels, which bind to syntaxin, SNAP-25, and synaptotagmin. Calcium has a biphasic effect on the interactions of N-type calcium channels with SNARE complexes, stimulating optimal binding in the range of 10-20 microM. PKC or CaM KII phosphorylation of the N-type synprint peptide inhibits interactions with native brain SNARE complexes containing syntaxin and SNAP-25. Introduction of the synprint peptides into presynaptic superior cervical ganglion neurons reversibly inhibits EPSPs from synchronous transmitter release by 42%. At physiological Ca2+ concentrations, synprint peptides cause an approximate 25% reduction in transmitter release of injected frog neuromuscular junction in cultures, consistent with detachment of 70% of the docked vesicles from calcium channels based on a theoretical model. Together, these studies suggest that presynaptic calcium channels not only provide the calcium signal required by the exocytotic machinery, but also contain structural elements that are integral to vesicle docking, priming, and fusion processes.
Sheng
Z H
ZH
Synaptic Function Unit, NINDS, NIH, Bethesda, MD 20892, USA.
Westenbroek
R E
RE
Catterall
W A
WA
D000200:Action Potentials / Q000187:drug effects; D000818:Animals; D001665:Binding Sites; D002121:Calcium Channel Blockers / Q000494:pharmacology; D015220:Calcium Channels / Q000187:drug effects / Q000502:physiology* / Q000648:ultrastructure; D020864:Calcium Channels, N-Type; D020013:Calcium Signaling; D002135:Calcium-Binding Proteins; D002478:Cells, Cultured; D006801:Humans; D046911:Macromolecular Substances; D008562:Membrane Glycoproteins / Q000502:physiology; D008565:Membrane Proteins / Q000502:physiology; D008958:Models, Molecular; D009419:Nerve Tissue Proteins / Q000187:drug effects / Q000502:physiology* / Q000648:ultrastructure; D018377:Neurotransmitter Agents / Q000378:metabolism; D017729:Presynaptic Terminals / Q000187:drug effects / Q000502:physiology* / Q000648:ultrastructure; D050765:Qa-SNARE Proteins; D051381:Rats; D050600:SNARE Proteins; D013329:Structure-Activity Relationship; D017783:Superior Cervical Ganglion / Q000166:cytology; D009435:Synaptic Transmission / Q000187:drug effects / Q000502:physiology*; D013572:Synaptic Vesicles / Q000187:drug effects / Q000502:physiology* / Q000648:ultrastructure; D050825:Synaptosomal-Associated Protein 25; D050857:Synaptotagmins; D033921:Vesicular Transport Proteins
D002121:Calcium Channel Blockers; D015220:Calcium Channels; D020864:Calcium Channels, N-Type; D002135:Calcium-Binding Proteins; D046911:Macromolecular Substances; D008562:Membrane Glycoproteins; D008565:Membrane Proteins; D009419:Nerve Tissue Proteins; D018377:Neurotransmitter Agents; D050765:Qa-SNARE Proteins; D050600:SNARE Proteins; D050825:Synaptosomal-Associated Protein 25; D050857:Synaptotagmins; D033921:Vesicular Transport Proteins; C494208:SNAP25 protein, human; C494210:Snap25 protein, rat