Ionotropic GABA receptors are widely distributed throughout the vertebrate and invertebrate central nervous system (CNS) where they mediate inhibitory neurotransmission. One of the most widely studied insect GABA receptors is constructed from RDL (resistance to dieldrin) subunits from Drosophila melanogaster. The aim of this study was to determine critical features of agonists binding to RDL receptors using in silico and experimental data. Partial atomic charges and dipole separation distances of a range of GABA analogues were calculated, and the potency of the analogues was determined using RDL receptors expressed in Xenopus oocytes. These data revealed functional agonists require an ammonium group and an acidic group with an optimum separation distance of approximately 5 A. To determine how the agonists bind to the receptor, a homology model of the extracellular domain was generated and agonists were docked into the binding site. The docking studies support the requirements for functional agonists and also revealed a range of potential interactions with binding site residues, including hydrogen bonds and cation-pi interactions. We conclude that the model and docking procedures yield a good model of the insect GABA receptor binding site and the location of agonists within it.