PHYSICAL LINK AND FUNCTIONAL COUPLING OF PRESYNAPTIC CALCIUM CHANNELSAND THE SYNAPTIC VESICLE DOCKING FUSION MACHINERY/

N- and P/Q-type calcium channels are localized in high density in pres ynaptic nerve terminals and are crucial elements in neuronal excitatio n-secretion coupling. In addition to mediating Ca2+ entry to initiate transmitter release, they are thought to interact directly with protei ns of the synaptic vesicle docking/fusion machinery. As outlined in th e 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 t he role of the synaptic protein interaction (synprint) sites in the in tracellular loop II-III (LII-III) of both alpha(1B) and alpha(1A) subu nits of N-type and P/Q-type calcium channels, which bind to syntaxin, SNAP-25, and synaptotagmin. Calcium has a biphasic effect on the inter actions of N-type calcium channels with SNARE complexes, stimulating o ptimal binding in the range of 10-20 mu M. PKC or CaM KII phosphorylat ion of the N-type synprint peptide inhibits interactions with native b rain SNARE complexes containing syntaxin and SNAP-25. Introduction of the synprint peptides into presynaptic superior cervical ganglion neur ons 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 neur omuscular junction in cultures, consistent with detachment of 70% of t he 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 doc king, priming, and fusion processes.