TY - JOUR
T1 - A rapid ion-exchange assay for detergent-solubilized inositol 1,4,5-trisphosphate receptors
AU - Hingorani, S. R.
AU - Agnew, William S.
PY - 1991/4
Y1 - 1991/4
N2 - We describe a rapid ion-exchange syringe assay for [3H]inositol 1,4,5-trisphosphate binding to detergent-solubilized receptors. In extracts of rat cerebellar membranes, the assay resolves rapidly dissociating ligand complexes, detecting two to three times higher receptor abundance than conventional gel filtration spun column assays, and provides evidence for two classes of IP3-binding sites, representing 0.5-1.0% of total cerebellar membrane protein. Receptors purified from bovine and rat cerebellum exhibit a single class of high-affinity sites, with equilibrium dissociation constants (Kd = 4-8 nM) reflecting 20 to 25-fold higher affinity than reported in studies with spun-column methods.
AB - We describe a rapid ion-exchange syringe assay for [3H]inositol 1,4,5-trisphosphate binding to detergent-solubilized receptors. In extracts of rat cerebellar membranes, the assay resolves rapidly dissociating ligand complexes, detecting two to three times higher receptor abundance than conventional gel filtration spun column assays, and provides evidence for two classes of IP3-binding sites, representing 0.5-1.0% of total cerebellar membrane protein. Receptors purified from bovine and rat cerebellum exhibit a single class of high-affinity sites, with equilibrium dissociation constants (Kd = 4-8 nM) reflecting 20 to 25-fold higher affinity than reported in studies with spun-column methods.
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U2 - 10.1016/0003-2697(91)90169-T
DO - 10.1016/0003-2697(91)90169-T
M3 - Article
C2 - 1651064
AN - SCOPUS:0025906212
SN - 0003-2697
VL - 194
SP - 204
EP - 213
JO - Analytical biochemistry
JF - Analytical biochemistry
IS - 1
ER -