对壳梭孢菌素响应的质子转运系统的功能重建。
Functional reconstitution of a proton-translocating system responsive to fusicoccin.
作者信息
Aducci P, Ballio A, Blein J P, Fullone M R, Rossignol M, Scalla R
机构信息
Dipartimento di Biologia, Università di Roma Tor Vergata, Italy.
出版信息
Proc Natl Acad Sci U S A. 1988 Nov;85(21):7849-51. doi: 10.1073/pnas.85.21.7849.
Abstract
Crude fusicoccin binding proteins and a partially purified plasma membrane H+-transporting ATPase (EC 3.6.1.34), both solubilized from maize tissues, were simultaneously inserted into liposomes by the freeze-thaw method. ATP-driven intravesicular acidification in the proteoliposomes, measured by the fluorescence quenching of the dye 9-amino-6-chloro-2-methoxyacridine, markedly increased upon addition of fusicoccin to the reconstituted system. This effect could not be observed when binding sites and ATPase preparations were separately reconstituted into the proteoliposomes, thus demonstrating that fusicoccin binding to its receptor is a prerequisite for ATPase stimulation.
摘要
从玉米组织中溶解得到的粗制壳梭孢菌素结合蛋白和部分纯化的质膜H⁺转运ATP酶（EC 3.6.1.34），通过冻融法同时插入脂质体中。通过染料9-氨基-6-氯-2-甲氧基吖啶的荧光猝灭来测量蛋白脂质体中ATP驱动的囊泡内酸化，当向重组系统中添加壳梭孢菌素时，这种酸化显著增加。当结合位点和ATP酶制剂分别重组到蛋白脂质体中时，未观察到这种效应，从而表明壳梭孢菌素与其受体的结合是ATP酶刺激的先决条件。
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本文引用的文献
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H-pumping driven by the plasma membrane ATPase in membrane vesicles from radish: stimulation by fusicoccin.H 泵在萝卜膜囊泡中的质膜 ATP 酶驱动下的作用： fusicoccin 的刺激作用。
Plant Physiol. 1986 Sep;82(1):121-5. doi: 10.1104/pp.82.1.121.
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The structure of fusicoccin A.壳梭孢菌素A的结构。
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Reconstitution, a way of biochemical research; some new approaches to membrane-bound enzymes.重组——一种生化研究方法；膜结合酶的一些新研究途径。
Arch Biochem Biophys. 1979 Dec;198(2):470-7. doi: 10.1016/0003-9861(79)90521-6.
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