Abstract

Calcium modulates sodium-independent binding of gamma-aminobutyric acid (GABA) to GABAA and GABAB recognition sites located in synaptic membranes of rat brain. At 37 degrees the binding of [3H]GABA to the GABAB recognition site is dramatically stimulated by Ca2+ with an EC50 (half-saturation constant) of congruent to 10 microns, whereas the binding to the GABAA recognition site is only slightly, but significantly, potentiated by Ca2+ with an EC50 of congruent to 0.1-1.0 micron. The effect of calcium on GABAA recognition sites requires a temperature of 37 degrees and the presence of calmodulin. Only GABAA recognition sites are linked to benzodiazepine recognition sites, and the interaction between these sites is modulated by Ca2+ at physiological ion concentrations. When free Ca2+ in the assay medium is below 10 nM, only one population of low-affinity GABAA recognition sites can be measured; however, when free Ca2+ is at the micromolar level, or if diazepam is present, a high affinity-binding site appears in addition to the pre-existing low-affinity component. On the basis of affinity there is a single population of GABAB bindings sites, but the number of sites is about 90% greater at 37 degrees than at 4 degrees. This temperature-dependent increase in the number of GABAB recognition sites is calmodulin-independent, and data with leupeptin, hemin, and antipain suggest that this temperature-dependent increase in GABAB sites might involve the activity of Ca2+-dependent protease(s).