CHARACTERIZATION AND POSTULATED STRUCTURE OF THE PRIMARY EMITTER IN THE BACTERIAL LUCIFERASE REACTION
CHARACTERIZATION AND POSTULATED STRUCTURE OF THE PRIMARY EMITTER IN THE BACTERIAL LUCIFERASE REACTION
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DOI：
10.1073/pnas.81.10.2990
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发表时间：
1984-01-01
期刊：
影响因子：
--
通讯作者：
HASTINGS, JW
中科院分区：
文献类型：
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作者：
KURFURST, M;GHISLA, S;HASTINGS, JW
一种可被鉴定为细菌（哈维氏弧菌）生物发光中的发射体的中间体被证实。该反应在1℃下进行，通过将纯化的与荧光素酶结合的黄素单核苷酸（FMN）4α - 氢过氧化物与长链醛（癸醛）混合。对生物发光和吸光度的同时动力学测量表明，发光的衰减比稳定产物氧化型FMN的出现更快，这表明在发光之后形成了一种瞬时中间物质。在发光完成后立即检测的反应混合物中发现了相同的物质。它具有相对较短的半衰期（在9℃下为7分钟）；发色团被假定为与荧光素酶结合的黄素4α - 氢氧化物，并通过失水衰变为稳定产物FMN。它的吸收光谱（λmax，360 nm）和它的荧光发射（λmax，490 nm）与这是初级发射体（反应中产生的生物发光物质）的基态这一假设相符。
An intermediate identifiable as the emitter in bacterial [Vibrio harveyi] bioluminescence was demonstrated. The reaction was carried out at 1.degree. C by mixing purified luciferase-bound FMN 4.alpha.-hydroperoxide with long-chain aldehyde (decanal). Simultaneous kinetic measurements of bioluminescence and absorbance showed that the decay of light emission occurred more rapidly than the appearance of the stable product, oxidized FMN, indicating the formation of a transient intermediate species subsequent to light emission. The same species was found in reaction mixtures examined immediately after light emission was completed. It has a relatively short half-life (7 min at 9.degree. C); the chromophore is postulated to be the luciferase-bound flavin 4.alpha.-hydroxide and to decay to the stable product, FMN, by losing water. Its absorption spectrum (.lambda.max, 360 nm) and its fluorescence emission (.lambda.max, 490 nm) are consistent with the hypothesis that this is the ground state of the primary emitter, the bioluminescent species produced in the reaction.