Quinoprotein Ethanol Dehydrogenase: Preparation of the Apo-form and Reconstitution with Pyrroloquinoline Quinone and Ca2+ or Sr2+ Ions
Quinoprotein Ethanol Dehydrogenase: Preparation of the Apo-form and Reconstitution with Pyrroloquinoline Quinone and Ca2+ or Sr2+ Ions
复制标题
喹啉蛋白乙醇脱氢酶：Apo 形式的制备和用吡咯喹啉醌和 Ca2 或 Sr2 离子重建
DOI：
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发表时间：
1991
期刊：
影响因子：
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通讯作者：
H. Görisch
中科院分区：
文献类型：
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作者：
A. Mutzel;H. Görisch
来自铜绿假单胞菌ATCC 17933的均质醌蛋白乙醇脱氢酶在天然酶的每个亚基中含有一个Ca 2+离子。在30°C下用反式-1，2-二氨基环己烷-N，N，N，N′N′-四乙酸处理产生催化非活性的脱辅基形式。在与Ca 2+和吡咯并喹啉醌孵育脱辅基形式后，重构完全活性的全酶。该重构酶的吸收光谱与天然酶的吸收光谱相同。脱辅基酶与Sr 2+和PQQ的孵育导致活性Sr 2+形式的形成。Sr 2+和Ca 2 +-形式的酶在其吸收光谱上不同。反式-1，2-二氨基环己烷-N，N，N，N′N′-四乙酸对Sr ~（2+）的灭活速度比Ca ~（2+）快一倍。Ca 2+是吡咯喹啉醌与醌蛋白乙醇脱氢酶的脱辅基形式结合所必需的。
Homogeneous quinoprotein ethanol dehydrogenase from Pseudomonas aeruginosa ATCC 17933 contains one Ca2+ ion per subunit of native enzyme. Treatment with fra/is-1,2-diaminocyclohexane-N,N,N,N′N′-tetraacetic acid at 30°C led to an catalytically inactive apo-form. Upon incubation of the apo-form with Ca2+ and pyrroloquinoline quinone a fully active holo-enzyme was reconstituted. The absorption spectrum of this reconstituted enzyme is identical to that of the native enzyme. Incubation of apo-enzyme with Sr2+ and PQQ led to the formation of an active Sr2+-form. The Sr2+ and the Ca2+-forms of the enzyme differ in their absorption spectra. The Sr2+-form was inactivated by frawv-1,2-diaminocyclohexane-N,N,N,N′N′-tetraacetic acid twice as fast as the Ca2+-form. Ca2+ is necessary for pyrroloquinoline quinone to bind to the apo-form of quinoprotein ethanol dehydrogenase.