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苹果果肉质膜微囊主动运输Ca2 的Ca2

周 卫 何 萍

摘要:应用45Ca2+示踪法研究了苹果果肉质膜微囊依赖于Ca2+的ATP酶(Ca2+-ATP酶)活性与Ca2+运输之间的关系及激素对该酶活性的影响。结果表明:Ca2+-ATP酶存在于质膜上并受载体A23187刺激而活性增加,该酶活性与依赖于ATP的Ca2+运输依抑制剂EB、游离Ca2+和ATP浓度的变化并呈极为相似的饱和动力学特征;而其EB半抑制浓度,Ca2+和ATP半饱和浓度分别为0.1,0.1和 50μmol/L,从而证实了正是Ca2+-ATP酶推动苹果果肉质膜微囊的Ca2+的主动运输。生长素与萘乙酸均可促进苹果果肉质膜微囊Ca2+-ATP酶活性和Ca2+吸收,而赤霉素则无此作用。

关键词:质膜微囊,Ca2+-ATP酶,Ca2+主动运输,激素,苹果果肉
学科分类号:Q946,Q942

Characterization of Ca2+-ATPase Which Drives Active Transport of Ca2+ in Plasma Membrane Vesicles Isolated from Pulp Cell of Malus pumila Fruits

ZHOU Wei and HE Ping
(Soil and Fertilizer Institute, Chinese Academy of Agricultural Sciences, Beijing 100081)

Abstract:The relationship between Ca2+-dependent ATPase(Ca2+-ATPase)and ATP-dependent Ca2+ transport in plasma membrane vesicles isolated from pulp cell of Malus pumila fruits and the influence of hormones on this enzyme were investigated by using 45Ca2+ tracing. Results showed that the Ca2+-ATPase is stimulated by the Ca2+ ionophore A23187(Fig.3) and is localized at the plasma membrane(Fig.1;Tables 1,2). Ca2+-ATPase activity and ATP-dependent Ca2+ uptake show very similar saturation kinetics with erythrosin B(50% inhibition at about 0.1μmol/L)(Fig.4),free Ca2+(half-maximal rate at about 0.1μmol/L)(Fig.5),and ATP(Km 50μmol/L)(Fig.2),demonstrating that it is this enzyme that drives ATP-dependent Ca2+ transport in plasma membrane vesicles. Both Ca2+-ATPase and Ca2+ uptake could be promoted also by IAA or NAA,but none by GA(Fig.6).

Key words:plasma membrane vesicles,Ca2+-ATPase,Ca2+ transport,hormone, pulp cell of Malus pumila fruits

   Ca2+能直接参与调节植物细胞内许多重要生理生化过程(周卫和林葆1995a,1995b,1996;Huang等1996;Paul等1996)。正常条件下,陡电化学势梯度有利于胞外Ca2+直接进入胞质(Donald 1990),但胞质内Ca2+必须保持极低的浓度(10-7~10-6mol/L)。胞质的低Ca2+浓度主要通过胞质Ca2+跨液泡膜、线粒体膜及内质网膜等膜系统主动运输进入细胞器,或由跨质膜主动运输进入胞外空间而实现的(Hepler和Wayne 1985)。研究表明,Ca2+跨膜运输包括初级主动运输和次级主动运输两方面。初级主动运输可由结合在内质网膜上依赖于Mg-ATP的Ca2+吸收系统(Bush等1986)和结

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