TY - JOUR T1 - Localization of synaptotagmin-binding domains on syntaxin JF - The Journal of Neuroscience JO - J. Neurosci. SP - 1975 LP - 1981 DO - 10.1523/JNEUROSCI.16-06-01975.1996 VL - 16 IS - 6 AU - Y Kee AU - RH Scheller Y1 - 1996/03/15 UR - http://www.jneurosci.org/content/16/6/1975.abstract N2 - Synaptotagmin, an abundant calcium- and phospholipid-binding protein of synaptic vesicles, has been proposed to regulate neurotransmitter release at the nerve terminal. To understand better the biochemical mechanism of neurotransmitter release, we have investigated the calcium- dependent and -independent protein-protein interactions between synaptotagmin I and syntaxin 1a, a subunit of the receptor for synaptic vesicles on the presynaptic plasma membrane. Soluble syntaxin 1a binds to synaptotagmin glutathione S-transferase (GST) fusion protein, and the binding was decreased in the presence of calcium. A synaptotagmin fragment containing the second C2 repeat (Syt3–5) had the same binding profile as the whole cytoplasmic domain; however, fragments containing the first C2 repeat (Syt1–3 and Syt2–3) showed calcium-dependent binding to syntaxin. In addition, the soluble full-length cytoplasmic domain of synaptotagmin binds to a syntaxin GST fusion protein in a calcium-dependent manner. Syntaxin domains required for calcium- dependent and -independent synaptotagmin-binding were localized using syntaxin deletion mutants. Amino acids 241–266 of the syntaxin C terminus were required for calcium-independent binding of synaptotagmin. The minimal domain required for calcium-dependent binding of synaptotagmin to syntaxin was localized to amino acids 220– 266. The syntaxin domains required for synaptotagmin binding overlap with the domains for vesicle-associated membrane protein (or VAMP) and alpha-soluble N-ethyl-maleimide-sensitive fusion protein attachment protein (or alphaSNAP) interactions. The data suggest both calcium- dependent and -independent roles of synaptotagmin in regulating synaptic vesicle release and/or recycling. ER -