通过使用标记前体推断血小板中肌醇脂质的亚细胞定位。
Subcellular localization of inositol lipids in blood platelets as deduced from the use of labelled precursors.
作者信息
Mauco G, Dajeans P, Chap H, Douste-Blazy L
机构信息
INSERM Unité 101, Hôpital Purpan, Toulouse, France.
出版信息
Biochem J. 1987 Jun 15;244(3):757-61. doi: 10.1042/bj2440757.
Abstract
- By rapid fractionation of blood platelet lysates on Percoll density gradients at alkaline pH (9.6), a very pure plasma-membrane fraction was obtained, as well as discrimination between endoplasmic reticulum and lysosomes. 2. Labelling of intact platelets with [32P]Pi followed by subcellular fractionation showed an exclusive localization of all inositol lipids in the plasma membrane. 3. Preincubation of whole platelets with myo-[3H]inositol in a buffer containing 1 mM-MnCl2 allowed incorporation of the label into PtdIns (phosphatidylinositol) of both plasma and endoplasmic-reticulum membrane, whereas [3H]PtdIns4P (phosphatidylinositol 4-phosphate) and [3H]PtdIns(4,5)P2 (phosphatidylinositol 4,5-bisphosphate) were exclusively found on the plasma membrane. 4. It is concluded that PtdIns4P and PtdIns(4,5)P2 are exclusively localized in the plasma membrane, whereas PtdIns is present in both plasma and endoplasmic-reticulum membranes. This could provide an explanation for previously reported data on hormone-sensitive and -insensitive inositol lipid pools.
摘要
- 通过在碱性pH值（9.6）下对血小板裂解物进行Percoll密度梯度快速分级分离，获得了非常纯净的质膜组分，并区分了内质网和溶酶体。2. 用[32P]Pi标记完整血小板，随后进行亚细胞分级分离，结果显示所有肌醇脂质都仅定位于质膜中。3. 在含有1 mM - MnCl2的缓冲液中，用肌醇-[3H]肌醇对全血小板进行预孵育，可使标记物掺入质膜和内质网膜的磷脂酰肌醇（PtdIns）中，而[3H]磷脂酰肌醇4-磷酸（PtdIns4P）和[3H]磷脂酰肌醇4,5-二磷酸（PtdIns(4,5)P2）仅存在于质膜上。4. 得出的结论是，PtdIns4P和PtdIns(4,5)P2仅定位于质膜中，而PtdIns存在于质膜和内质网膜中。这可以为先前报道的关于激素敏感和不敏感肌醇脂质池的数据提供解释。
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