A novel, specific binding protein assay for quantitation of intracellular inositol 1,3,4,5-tetrakisphosphate (InsP4 ) using a high-affinity InsP4 receptor from cerebellum
- 28 August 1989
- journal article
- Published by Wiley in FEBS Letters
- Vol. 254 (1-2) , 155-158
- https://doi.org/10.1016/0014-5793(89)81029-4
Abstract
A membrane preparation from porcine cerebellum displays high-affinity binding sites for [3H]inositol 1,3,4,5-tetrakisphosphate ([3H]InsP4) with a dissociation constant (K d) of 1.0 nM and a density of 220 fmol/mg protein. Specific binding was maximal in the presence of 25 mM phosphate and at pH 5.0. The receptor site was specific for InsP4, since Ins(1,3,4,5,6)P5 and Ins(1,4,5,6)P4 displaced binding of InsP4 with EC50 values of 0.2 and 0.3 μM, respectively. Ins(1,4,5)P3 and other inositol phosphates were less effective. Using this InsP4 receptor, an assay for measuring tissue content of InsP4 was developed. The detection limit of the assay was 0.1 pmol. In the same tissue samples the amount of Ins(1,4,5)P3 was determined in parallel with a similar assay using a binding protein preparation from beef liver.Keywords
This publication has 14 references indexed in Scilit:
- Development of a novel, Ins(1,4,5)P3-specific binding assay: Its use to determine the intracellular concentration of Ins(1,4,5)P3 in unstimulated and vasopressin-stimulated rat hepatocytesCellular Signalling, 1989
- A simple, sensitive, and specific radioreceptor assay for inositol 1,4,5-trisphosphate in biological tissuesBiochemical and Biophysical Research Communications, 1989
- Distinct binding sites for Ins(1,4,5)P3 and Ins(1,3,4,5)P4 in bovine parathyroid glandsBiochemical and Biophysical Research Communications, 1989
- Mass measurements of inositol(1,4,5)trisphosphate in rat cerebral cortex slices using a radioreceptor assay: Effects of neurotransmitters and depolarizationBiochemical and Biophysical Research Communications, 1988
- Inositol 1,3,4,5-Tetrakisphosphate Induces Ca 2+ Sequestration in Rat Liver CellsScience, 1988
- Specific binding sites for [3H]inositol(1,3,4,5)tetrakisphosphate on membranes of HL-60 cellsBiochemical and Biophysical Research Communications, 1987
- The inositol tris/tetrakisphosphate pathway—demonstration of Ins(l,4,5)P3 3-kinase activity in animal tissuesNature, 1986
- Bradykinin‐induced transient accumulation of inositol trisphosphate in neuron‐like cell line NG 108‐15 cellsFEBS Letters, 1985
- [26] Culture and characteristics of hormone-responsive neuroblastoma x glioma hybrid cellsPublished by Elsevier ,1985
- Inositol trisphosphate, a novel second messenger in cellular signal transductionNature, 1984