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Updated: Aug 23, 2026

Direct Imaging of ER Calcium with Targeted-Esterase Induced Dye Loading (TED)
Published on: May 7, 2013
Heterotrimeric Gi protein is associated with the inositol 1,4,5-trisphosphate receptor complex and modulates calcium
C B Neylon1, A Nickashin, V A Tkachuk
1Baker Medical Research Institute, Prahran, Victoria, Australia. neylon@neuro.duke.edu
Abstract:
In vascular smooth muscle, pertussis toxin (PT) inhibits thrombin-induced Ca2+ release by a mechanism independent of its effect on IP3 formation. Thus, the possibility of a direct role of G alpha i proteins in regulating IP3-sensitive Ca2+ release was investigated by examining whether G alpha i proteins are associated with the IP3 receptor complex. Purified microsomal membranes were prepared and separated by sucrose density gradient centrifugation. The relative density of [3H]-IP3 binding sites between the microsomal fractions was inversely related to the distribution of the plasma membrane marker. The relative distribution of G alpha i3 determined by immunoblotting was closely correlated with the density of [3H]-IP3 binding. Levels of G alpha i2 were more evenly distributed with highest levels present in plasma membrane-enriched fractions. IP3 receptor immunoprecipitated from triton-solubilized microsomal membranes contained G alpha i3 immunoreactivity. To determine whether G alpha i proteins influence IP3-induced Ca2+ release, the effect of PT on Ca2+ release from digitonin-permeabilized cell suspensions using Fluo-3 was examined. Exposure to PT (0.1 microgram/ml, 5 min) attenuated the initial rate of IP3 (1 microM)-induced Ca2+ release. Together, these findings are consistent with the hypothesis that a heterotrimeric G alpha i protein directly regulates IP3-dependent Ca2+ release.
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