Body plasma membrane vesicles from Paramecium contain a vanadate-sensitive Ca(2+)-ATPase.
Body plasma membrane vesicles from Paramecium contain a vanadate-sensitive Ca(2+)-ATPase.
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草履虫体内质膜囊泡含有对钒酸盐敏感的 Ca(2)-ATP 酶。
DOI：
10.1006/abio.1993.1199
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发表时间：
1993
影响因子：
2.9
通讯作者：
Hennessey,TM
中科院分区：
文献类型：
--
作者：
Smith,TM;Hennessey,TM
Paramatrium是一个很好的模型系统，用于研究参与感觉转导和细胞内Ca 2+调节的机制。这些细胞具有两个功能不同的质膜结构域，体和纤毛。身体质膜负责将感觉刺激转导成受体电位，并且睫状膜是Ca 2+动作电位所需的。虽然睫状膜囊泡（cmv）已被纯化和良好的特点，身体质膜还没有。我们通过匀浆去纤毛细胞产生了体质膜囊泡，并通过两相水相聚合物分离从微粒体组分中纯化了体质膜囊泡。bmv级分纯度的主要标准是：（i）已知质膜标记物（固定抗原）富集15倍，而其他膜的标记物活性均降低。bmv的蛋白质电泳带型与cmv相似。(ii)它含有钒酸盐敏感的Ca 2 +-ATP酶活性，被认为是质膜Ca 2+泵。此bmv Ca ~（2+）-ATP酶的比活性比匀浆的比活性增加4倍。(iii)bmv组分中的磷脂、脂肪酸和甾醇成分是质膜的指示，因为它们在性质上与cmv相似。bmv还含有膜结合的NADPH依赖性细胞色素c还原酶活性，这表明它可能在身体质膜功能中发挥作用。这种纯化的bmv制剂是有用的研究的作用，身体质膜在Ca 2+调节，感觉转导，蛋白质和脂质贩运，质膜融合事件。
Paramecia are an excellent model system for studying the mechanisms involved in sensory transductions and intracellular Ca2+regulation. These cells have two functionally distinct plasma membrane domains, body and cilia. The body plasma membrane is responsible for transduction of sensory stimuli into receptor potentials and the ciliary membrane is required for Ca2+action potentials. Although ciliary membrane vesicles (cmv) have been purified and well characterized, body plasma membranes have not. We have generated body plasma membrane vesicles (bmv) by homogenization of deciliated cells and purified them from the microsome fraction by a two-phase aqueous polymer separation. The major criteria for purity of the bmv fraction are: (i) It is enriched 15-fold for a known plasma membrane marker (immobilization antigen) while the marker activities for other membranes were all decreased. The protein banding pattern of bmv is generally similar to cmv on SDS-PAGE. (ii) It contains a vanadate-sensitive Ca2+-ATPase activity that has been suggested to be a plasma membrane Ca2+pump. The specific activity of this bmv Ca2+-ATPase is increased 4-fold over that of the homogenate. (iii) The phospholipid, fatty acid, and sterol composition of the bmv fraction are indicative of plasma membranes because they are qualitatively similar to cmv. The bmv also contains a membrane-bound NADPH-dependent cytochrome c reductase activity, suggesting that it may play a role in body plasma membrane function. This purified bmv preparation is useful for studying the role of the body plasma membrane in Ca2+regulation, sensory transduction, protein and lipid trafficking, and plasma membrane fusion events.