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MES [2-(N-Morpholino)ethanesulfonic Acid]

MES [2-(N-Morpholino)ethanesulfonic Acid]

Company: NACALAI TESQUE
Catalog#: 21623-26
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Measurement of ATP Hydrolytic Activity of Plasma Membrane H+-ATPase from Arabidopsis thaliana Leaves
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2016-12-05
[Abstract]  Plant plasma membrane H+-ATPase, which is a P-type ATPase, couples ATP hydrolysis to H+ extrusion and thereby generates an electrochemical gradient across the plasma membrane. The proton gradient is necessary for secondary transport, cell elongation, and membrane potential maintenance. Here we describe a protocol for measurement of the ATP hydrolytic activity of the plasma membrane H+-ATPase from Arabidopsis thaliana leaves. [摘要]  作为P型ATP酶的植物质膜H sup + -ATPase将ATP水解耦合到H + +/- 挤出,从而在质膜上产生电化学梯度。质子梯度对于二次转运,细胞伸长和膜电位维持是必需的。这里我们描述用于测量来自拟南芥叶片的质膜H + -ATPase的ATP水解活性的方案。
关键词: strong> 拟南芥,ATP水解活性,原钒酸盐,P-型ATP酶,血浆膜H + -ATPase

质膜H + -ATPase活性的测定对于阐明其功能和调节机制是重要的。然而,有时难以确定质膜H sup + -ATP酶的ATP水解活性,因为植物细胞含有许多ATP水解酶。该协议是基于Uemura和Yoshida(1986)和Kinoshita等人的出版物开发的。 (1995)。我们使用KNO 3作为V型ATP酶的抑制剂,钼酸铵作为酸性磷酸酶的抑制剂,寡霉素作为F型ATP酶的抑制剂,NaF作为磷酸酶的抑制剂(Shimazaki和Kondo ,1987; Kinoshita等人,1995)。原钒酸盐抑制P型ATP酶,并且因此可以通过评估来自ATP水解的钒酸盐敏感性P 1释放来用于测量质膜H sup + -ATP酶的活性。释放的P 1与钼酸盐反应形成蓝色络合物,然后可以通过测量在750nm的吸收来量化。

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