The cytomatrix at the active zone (CAZ) has been implicated in defining the site of Ca2+-dependent exocytosis of neurotransmitter. We have identified here a novel CAZ protein of ∼120 kD from rat brain and named it CAST (CAZ-associated structural protein). CAST had no transmembrane segment, but had four coiled-coil domains and a putative COOH-terminal consensus motif for binding to PDZ domains. CAST was localized at the CAZ of conventional synapses of mouse brain. CAST bound directly RIM1 and indirectly Munc13-1, presumably through RIM1, forming a ternary complex. RIM1 and Munc13-1 are CAZ proteins implicated in Ca2+-dependent exocytosis of neurotansmitters. Bassoon, another CAZ protein, was also associated with this ternary complex. These results suggest that a network of protein–protein interactions among the CAZ proteins exists at the CAZ. At the early stages of synapse formation, CAST was expressed and partly colocalized with bassoon in the axon shaft and the growth cone. The vesicles immunoisolated by antibassoon antibody–coupled beads contained not only bassoon but also CAST and RIM1. These results suggest that these CAZ proteins are at least partly transported on the same vesicles during synapse formation.
Cast : a novel protein of the cytomatrix at the active zone of synapses that forms a ternary complex with RIM1 and munc13-1
Abbreviations used in this paper: APMSF, α-amidinophenyl-methanesulfonyl fluoride hydrochloride; CAST, CAZ-associated structural protein; CAZ, cytomatrix at the active zone; P2, crude membrane; P100, pellet; pBS, pBluescript-SK; PSD, postsynaptic density; S100, supernatant; SAP, synapse-associated protein.
Toshihisa Ohtsuka, Etsuko Takao-Rikitsu, Eiji Inoue, Marie Inoue, Masakazu Takeuchi, Kaho Matsubara, Maki Deguchi-Tawarada, Keiko Satoh, Koji Morimoto, Hiroyuki Nakanishi, Yoshimi Takai; Cast : a novel protein of the cytomatrix at the active zone of synapses that forms a ternary complex with RIM1 and munc13-1 . J Cell Biol 5 August 2002; 158 (3): 577–590. doi: https://doi.org/10.1083/jcb.200202083
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