Intrinsic inhibitor of inositol 1,4,5-trisphosphate binding |
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Authors: | Masato Hirata Masako Yoshida Takashida Kanematsu Hiroshi Takeuchi |
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Institution: | (1) Department of Biochemistry, Faculty of Dentistry, Kyushu University, Fukuoka, Japan |
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Abstract: | Rat brain cytosol was applied to a heparin column and eluted with 0.9 M-NaCl. The total binding activity of 3H]inositol 1,4,5-trisphosphate to the eluate was increased about 6-fold compared with the original cytosol. When the eluate was mixed with a flow-through fraction from the heparin column, however, the activity returned to the original level, suggesting that the flow-through fraction contained an inhibitory factor(s) which prevented the binding. The factor(s) was purified by sequential column chromatography using gel permeation, a hydrophobic gel, and finally, a hydroxylapatite gel. Silver staining of sodium dedecyl sulfate gel electrophoresis of the sample thus purified showed a broad band located between the authentic molecular weight markers of 580 and 390 k. A carbohydrate staining method showed that the factor is a glycoprotein. |
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Keywords: | inositol 1 4 5-triphosphate receptor pleckstnin homology domain |
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