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- W985333010 abstract "This chapter presents protocol for the purification of novel Ins (l, 4, 5) P3-binding proteins from rat brain cytosol, using the Ins (l, 4, 5) P 3 affinity matrices. Myo-inositol 1,4,5-trisphosphate [Ins(l,4,5)P3 ], a product of the receptor-activated hydrolysis of phosphatidylinositol 4,5-bisphosphate (PIP2 ), plays an important role as an intracellular second messenger by mobilizing Ca2+ from nonmitochondrial store sites. The analog of Ins (l, 4, 5) P3 has the azidobenzoyl group at the C-2 position for photoaffinity labeling and causes irreversible inactivation of the receptor protein for Ca2+ release following photolysis. The biological activities of Ins (l, 4, 5) P3 are related to two adjacent phosphates at C-4 and C-5 and the phosphate at C-1 that increases the affinity for its recognition by the receptor site. The Ins (l, 4, 5) P3 analogs attach to other molecules through substituents at the C-2 position. The purification was most satisfactory with Ins (l, 4, 5) P3 affinity chromatography and the subsequent gel-filtration chromatography that separated the two binding activities with moderate purification. These findings suggest that the crude cytosol fraction have contained inhibitory factor(s) removable by application to an affinity column. Subsequent anion-exchange chromatography led to further purification, as assessed by SDS-polyacrylamide gel electrophoresis." @default.
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- W985333010 date "1993-01-01" @default.
- W985333010 modified "2023-10-15" @default.
- W985333010 title "Inositol 1,4,5-Trisphosphate-Binding Proteins in Rat Brain Cytosol" @default.
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- W985333010 doi "https://doi.org/10.1016/b978-0-12-185285-6.50031-2" @default.
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