通过黄嘌呤/黄嘌呤氧化酶法测定蛋白质中的黄素电位
Determination of Flavin Potential in Proteins by Xanthine/Xanthine Oxidase Method.
作者信息
Maklashina Elena, Cecchini Gary
机构信息
Molecular Biology Division, San Francisco VA Health Care System, San Francisco, CA 94121, USA.
Department of Biochemistry & Biophysics, University of California, San Francisco, Ca 95158, USA.
出版信息
Bio Protoc. 2020 Apr 5;10(7):e3571. doi: 10.21769/BioProtoc.3571.
Abstract
This protocol describes a simple xanthine/xanthine oxidase enzymatic equilibration method for determination of the redox potential of a flavin. As an example of the use of this method, we determine the reduction potential of the covalently bound FAD cofactor ( = -55 mV) in the SdhA flavoprotein subunit of succinate dehydrogenase from . In principle, this method can be used routinely to determine the redox potential of flavin cofactors in any simple flavoprotein from equilibrium concentrations with an appropriate reference dye of known without the use of sophisticated electrochemical equipment.
摘要
本方案描述了一种用于测定黄素氧化还原电位的简单黄嘌呤/黄嘌呤氧化酶酶促平衡方法。作为该方法应用的一个例子，我们测定了来自[具体来源]的琥珀酸脱氢酶SdhA黄素蛋白亚基中与共价结合的FAD辅因子的还原电位（=-55 mV）。原则上，该方法可常规用于通过与已知[具体参数]的合适参比染料达到平衡浓度来测定任何简单黄素蛋白中黄素辅因子的氧化还原电位，而无需使用复杂的电化学设备。
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