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Inhibitory effect of adenine nucleotides and anti‐allergic drugs on phosphorylation of phosphatidylinositol in rat mast cell granules
Author(s) -
KUROSAWA M.,
OKAYAMA Y.,
KOBAYASHI S.
Publication year - 1989
Publication title -
allergy
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 3.363
H-Index - 173
eISSN - 1398-9995
pISSN - 0105-4538
DOI - 10.1111/j.1398-9995.1989.tb04203.x
Subject(s) - phosphatidylinositol , chemistry , phosphorylation , percoll , mast cell , biochemistry , microbiology and biotechnology , theophylline , biology , pharmacology , in vitro , immunology
Rat mast cell granules were obtained by sonication of highly purified rat mast cells and isolated in a Percoll gradient. Phosphorylation of endogenous phosphatidylinositol in rat mast cell granules, which is catalyzed by phosphatidylinositol kinase in the granules, was assayed by measuring the incorporation of 32 P from [γ 32 P]ATP into phosphatidylinositol 4‐phosphate. Lipids were isolated with methanol/chloroform/HCl and were separated by thin‐layer chromatography on oxalic acid impregnated silica gel plates. Phosphatidylinositol 4‐phosphate areas were identified by staining with iodine, scraped and measured for 32 P radioactivity. The phosphorylation reaction was inhibited by 50–500 μM adenosine, ADP and 500 μM AMP in a concentration‐dependent manner. Among several antiallergic drugs investigated, 100±1000 μM theophylline and 10±100 μM azelastine inhibited the phosphorylation reaction, but disodium cromoglycate and ketotifen had littel effect.