Duine J A, Frank J, Verwiel P E
Eur J Biochem. 1980;108(1):187-92. doi: 10.1111/j.1432-1033.1980.tb04711.x.
The reconstructive ability of the isolated prosthetic group of methanol dehydrogenase with the apoenzyme of glucose dehydrogenase and the results of electron spin resonance measurements suggest that the prosthetic group has not been modified during the isolation. This result, and the properties of the directly isolated prosthetic group and derivatives, confirm the suggestion that its structure is 2,7,9-tricarboxy-1H-pyrrolo[2,3-f]quinoline-4,5-dione. From the activity shown by derivatives of the prosthetic group and of structural analogues in the apoenzyme test it is concluded that the o-quinone structure is essential for activity. Hence the trivial name pyrrolo-quinoline quinone would be appropriate. The testing of the analogues also shows that the pyrrolo ring and the 9-carboxylic acid group are not essential for activity as they can be replaced by a pyridinol ring and a 9-hydroxy group respectively. The determination of the molar absorption coefficient of the prosthetic group (18 400 M-1 cm-1 at 249 nm) enables its quantitative anaysis. Thus it could be established that methanol dehydrogenase contains one prosthetic group per enzyme molecule. The consequences of this result in relation to already known properties of this 'quinoprotein' dehydrogenase are discussed.
甲醇脱氢酶的分离辅基与葡萄糖脱氢酶脱辅基酶的重建能力以及电子自旋共振测量结果表明,该辅基在分离过程中未发生修饰。这一结果,以及直接分离得到的辅基及其衍生物的性质,证实了其结构为2,7,9 - 三羧基 - 1H - 吡咯并[2,3 - f]喹啉 - 4,5 - 二酮的推测。从辅基衍生物和结构类似物在脱辅基酶测试中所显示的活性可以得出结论,邻醌结构对于活性至关重要。因此,俗名吡咯并喹啉醌是合适的。对类似物的测试还表明,吡咯环和9 - 羧酸基团对于活性并非必不可少,因为它们可以分别被吡啶醇环和9 - 羟基取代。辅基摩尔吸收系数的测定(在249 nm处为18400 M-1 cm-1)使其能够进行定量分析。由此可以确定,甲醇脱氢酶每个酶分子含有一个辅基。讨论了这一结果与这种“醌蛋白”脱氢酶已知性质相关的后果。