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首页> 外文期刊>The protein journal >Different States of Acrylodan-Labeled 3'5'-Cyclic Adenosine Monophosphate Dependent Protein Kinase Catalytic Subunits in Denaturant Solutions
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Different States of Acrylodan-Labeled 3'5'-Cyclic Adenosine Monophosphate Dependent Protein Kinase Catalytic Subunits in Denaturant Solutions

机译：变性溶液中丙烯腈标记的3'5'-环磷酸腺苷依赖性蛋白激酶催化亚基的不同状态

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摘要

Fluorescence spectroscopy was used to differentiate between different states of acrylodan-labeled cAMP-dependent protein kinase catalytic subunits in urea, guanidine hydrochloride and 3-(N-morpholino)propanesulfonic acid solutions, by measuring changes in the emission spectrum of the protein-coupled dye, which is very sensitive to its microenvironment. Decomposition of the observed fluorescence spectra by a parameterized log-normal distribution function allowed the resolution of overlapping spectral bands and revealed the formation of three distinct protein states, denominated as native, denatured and unfolded structures. At low denaturant concentrations the formation of the denatured form from the native protein was observed, and this process was characterized by a blue-shift of the fluorescence spectrum of acrylodan, indicating that the dye was transferred into some water-deficit hydrophobic environment inside the protein molecule. Therefore, formation of a "dry molten globule" structure could be suggested in state. At high denaturant concentrations a red-shift of the emission spectrum of the protein-coupled probe was observed indicating significant extrusion of the dye molecule into water environment as a result of the unfolding of the protein structure.
机译：通过测量蛋白质偶联染料的发射光谱变化，使用荧光光谱法区分尿素，盐酸胍和3-（N-吗啉代）丙烷磺酸溶液中丙烯酰胺标记的cAMP依赖性蛋白激酶催化亚基的不同状态。 ，对其微环境非常敏感。通过参数化的对数正态分布函数对观察到的荧光光谱进行分解，可以分辨出重叠的光谱带，并揭示了三种不同的蛋白质状态的形成，分别称为天然，变性和未折叠结构。在低变性剂浓度下，观察到天然蛋白质会形成变性形式，并且该过程的特征是丙烯醛荧光光谱发生蓝移，这表明染料已转移到蛋白质内部某些缺水的疏水环境中分子。因此，可以建议在状态下形成“干燥熔融小球”结构。在高变性剂浓度下，观察到蛋白质偶联探针发射光谱的红移，表明由于蛋白质结构的展开，染料分子显着挤出到水环境中。

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