A simple, sensitive, and specific radioreceptor assay for inositol 1,4,5-trisphosphate in biological tissues

Inositol (1,4,5)P 3 (IP 3) receptors in rat cerebellar membranes have been used to develop a radioreceptor assay for endogenous IP 3. The assay is sensitive, detecting as little as 1 pmole IP 3 in a 0.5 ml volume. The receptor is highly selective for the 1,4,5-isomer of IP 3 so that assays can emplo...

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Veröffentlicht in:	Biochem. Biophys. Res. Commun.; (United States) 1989-03, Vol.159 (3), p.976-982
Hauptverfasser:	Bredt, David S., Mourey, Robert J., Snyder, Solomon H.
Format:	Artikel
Sprache:	eng
Schlagworte:	550201 - Biochemistry- Tracer Techniques Analytical, structural and metabolic biochemistry ANIMALS BASIC BIOLOGICAL SCIENCES Biological and medical sciences BODY BRAIN Calcium Channels CARBOHYDRATES CELL CONSTITUENTS Cell Membrane - metabolism CELL MEMBRANES CENTRAL NERVOUS SYSTEM CEREBELLUM Cerebellum - metabolism CHEMICAL REACTIONS CHROMATOGRAPHY Chromatography, Ion Exchange DECOMPOSITION ENZYMATIC HYDROLYSIS ESTERS Fundamental and applied biological sciences. Psychology HYDROLYSIS INOSITOL Inositol 1,4,5-Trisphosphate Inositol 1,4,5-Trisphosphate Receptors Inositol Phosphates - analysis Inositol Phosphates - isolation & purification Inositol Phosphates - metabolism INOSITOLS ION EXCHANGE CHROMATOGRAPHY ISOTOPE APPLICATIONS LIPIDS LYSIS Male MAMMALS MEMBRANE PROTEINS MEMBRANES MONOSACCHARIDES NERVOUS SYSTEM ORGANIC COMPOUNDS ORGANIC PHOSPHORUS COMPOUNDS ORGANS Other biological molecules PHOSPHOLIPIDS PROTEINS Radioligand Assay - methods RADIORECEPTOR ASSAY RATS Rats, Inbred Strains RECEPTORS Receptors, Cell Surface - metabolism Receptors, Cytoplasmic and Nuclear RODENTS SACCHARIDES SEPARATION PROCESSES SOLVOLYSIS Sugar Phosphates - analysis TRACER TECHNIQUES Tritium VERTEBRATES
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Zusammenfassung:	Inositol (1,4,5)P 3 (IP 3) receptors in rat cerebellar membranes have been used to develop a radioreceptor assay for endogenous IP 3. The assay is sensitive, detecting as little as 1 pmole IP 3 in a 0.5 ml volume. The receptor is highly selective for the 1,4,5-isomer of IP 3 so that assays can employ crude tissue extracts with no purification. Chromatographic analysis and selective enzymatic hydrolysis of IP 3 show that in crude tissue extracts only authentic IP 3 is detected. A hundred or more samples can be assayed in a day for endogenous IP 3.
ISSN:	0006-291X 1090-2104
DOI:	10.1016/0006-291X(89)92204-3