COLORS OF FIREFLY BIOLUMINESCENCE - ENZYME CONFIGURATION + SPECIES SPECIFICITY
COLORS OF FIREFLY BIOLUMINESCENCE - ENZYME CONFIGURATION + SPECIES SPECIFICITY
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DOI：
10.1073/pnas.52.1.75
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发表时间：
1964-01-01
影响因子：
11.1
通讯作者：
MCELROY, WD
中科院分区：
文献类型：
--
作者：
SELIGER, HH;MCELROY, WD
E+ LH 2 + ATP=-E。LH 2AMP + PP，（1）只有D（-）LH 2AMP将进一步反应，在氧气的存在下，产生生物发光和氧化产物。在pH< 6.5的酸性缓冲溶液中，正常的浅绿色发光强度（峰值在562 mU）显著降低，而观察到低强度的红色发光（峰值在616 mU）。这表明酶的构型在决定负责发光的激发态的共振能级方面是重要的。进一步证明民族的变化。除了酸性缓冲液中酶的部分变性外，pH对发射光谱漂移的影响是完全可逆的。我们已经能够观察到这些相同的可逆的红移发射光谱通过增加反应的温度，通过进行反应在0.2 M尿素和在正常pH值（7.6）的甘氨酰甘氨酸缓冲液中，通过添加小浓度的Zn++，Cd++，和Hg++阳离子，作为氯化物。不同Zn++浓度下纯化的萤火虫荧光素酶体外生物发光的归一化发射光谱见图1。的
E+ LH2+ ATP=-E. LH2AMP+ PP,(1) only D (-) LH2AMP will react further, in the presence of oxygen, to produce bioluminescence and an oxidized product. There is also a strong pH dependence of the color of the emitted light; 2 in acidic buffer solutions, pH< 6.5, the intensity of the normalyellow-green emission, peaking at 562 ml,, decreases markedly and a low intensity red emission is observed, peaking at 616 miu. This is evidence that enzyme configuration is important in determining theresonance energy levels of the excited state responsible for light emission. Further Evidence for Configurational Changes.-Except for the partial denaturation of the enzyme in acidic buffer, the pH effect on the emission spectrum shift is completely reversible. We have been able to observe these same reversible red shifts in emission spectra by increasing the temperature of the reaction, by carrying out the reaction in 0.2 M urea and at normal pH values (7.6) in glycyl glycine buffer, by adding small concentrations of Zn++, Cd++, and Hg++ cations, as chlorides. The normalized emission spectra of the in vitro bioluminescence of purified Photinus pyralis luciferase for various Zn++ concentrations are shown in Figure 1. The