The stoned locus in Drosophila encodes two proteins StonedA (STNA) and StonedB (STNB), both of which have been suggested to act as adaptins in mediating synaptic vesicle recycling. A combination of immunological, genetic and biochemical studies have shown an interaction of STNA and STNB with the C2B domain of Synaptotagmin‐I (SYT‐1), an integral synaptic vesicle protein that mediates Ca 2+ ‐dependent exocytosis, as well as endocytosis. The C2B domain of SYT‐1 contains an AP‐2 binding site that controls the size of recycled vesicles, and a C‐terminal tryptophan‐containing motif that acts as an internalization signal. Investigation of SYT‐1 mutations in Drosophila has shown that altering the Ca 2+ binding region of the C2B domain, results in a reduction in the rate of vesicle recycling, implicating this region in SYT‐I endocytosis. In this poster, we report the molecular dissection of the interactions between the STNA and STNB proteins and the C2B domain of SYT‐1. Deletion of the AP‐2 binding site decreased the binding of both STNA and STNB. However, C‐terminal deletions of the C2B domain significantly increased STNB binding. In contrast, the same C‐terminal deletions reduced the affinity of the C2B domain for STNA. The possible interactions of both STNB and STNA with the Ca 2+ binding region of SYT‐1 will be also investigated.