首页> 外文期刊>Proceedings of the Indian Academy of Sciences. Chemical Sciences >Time-resolved fluorescence study of the single tryptophan in thiocyanate and azide derivatives of horseradish peroxidase: Implication for a pH-induced conformational change in the heme cavity
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Time-resolved fluorescence study of the single tryptophan in thiocyanate and azide derivatives of horseradish peroxidase: Implication for a pH-induced conformational change in the heme cavity

机译:辣根过氧化物酶硫氰酸盐和叠氮化物衍生物中单个色氨酸的时间分辨荧光研究:对血红素腔中pH诱导的构象变化的影响

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摘要

Detailed pH-dependent steady state and picosecond time-resolved tryptophan fluorescence studies on thiocyanate and azide complexes of horseradish peroxidase have been carried out. The fluorescence decay of the single tryptophan in these species was fitted to a discrete three exponential model. Maximum entropy method analysis also gave three distinct regions of lifetime distributions. The fast subnanosecond lifetime component was found to have > 97% amplitude contribution while other two longer lifetime components have small contributions. Small contributions from the nanosecond lifetime components possibly arise from apo-protein impurity or some small amount of disordered heme conformer of the protein. pH dependence of the fast picosecond lifetime components was found to show a systematic behavior which has been interpreted in the light of obligatory conformation change associated with activation of the enzyme at low pH.
机译:已对辣根过氧化物酶的硫氰酸盐和叠氮化物复合物进行了详细的pH依赖性稳态和皮秒时间分辨色氨酸荧光研究。将这些物种中单个色氨酸的荧光衰减拟合为离散的三指数模型。最大熵方法分析还给出了寿命​​分布的三个不同区域。发现亚纳秒级快速寿命分量的振幅贡献大于97%,而其他两个较长寿命的分量贡献较小。纳秒寿命成分的少量贡献可能来自脱辅基蛋白杂质或少量的无序血红素构象异构体。已发现快速皮秒寿命组件的pH依赖性显示出系统的行为,该行为已根据与低pH值下酶的活化相关的强制性构象变化进行了解释。

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