Fluorescence and circular dichroism spectroscopy of cytochrome c in alkylammonium formate ionic liquids.
Fluorescence and circular dichroism spectroscopy of cytochrome c in alkylammonium formate ionic liquids.
复制标题
DOI：
10.1021/bm1008052
复制
发表时间：
2011-02-14
影响因子：
6.2
通讯作者：
Danielson, Neil D.
中科院分区：
文献类型：
--
作者：
Wei, Wenjun;Danielson, Neil D.
用荧光光谱和圆二色光谱研究了细胞色素c在甲酸烷基铵（AAF）离子液体（如甲酸甲铵（MAF）和甲酸乙铵（EAF））中的结构稳定性。在室温下，细胞色素c的天然结构在相对高的离子液体浓度（50%-70%AAF/水或AAF/磷酸盐缓冲液pH 7.0）下得以保持，这与细胞色素c在甲醇或乙腈的类似溶液中、在水或缓冲液共溶剂下的变性形成对比。荧光光谱和圆二色性光谱表明，在30 - 50 ℃的20%AAF-80%水中，细胞色素c的构象保持不变。在80%AAF-20%水中没有发现这种温度稳定性。与磷酸盐缓冲液相比，细胞色素c在80%AAF-20%水中的酶活性可以保持约三分之一，这大于在相应的甲醇和乙腈水溶液中测量的活性。该生物物理研究表明，AAF在中等温度下在移动的相中作为有机溶剂替代物具有潜在的应用，用于通过反相液相色谱法分离天然形式的蛋白质。
The structural stability of cytochrome c has been studied in alkylammonium formate (AAF) ionic liquids such as methylammonium formate (MAF) and ethylammonium formate (EAF) by fluorescence and circular dichroism (CD) spectroscopy. At room temperature, the native structure of cytochrome c is maintained in relatively high ionic liquid concentrations (50%–70% AAF/water or AAF/phosphate buffer pH 7.0) in contrast to denaturation of cytochrome c in similar solutions of methanol or acetonitrile, with water or buffer co-solvents. Fluorescence and CD spectra indicate the conformation of cytochrome c is maintained in 20% AAF-80% water from 30 – 50 °C. No such temperature stability is found in 80% AAF-20% water. About one third of the enzyme activity of cytochrome c in 80% AAF-20% water can be maintained as compared to phosphate buffer and this is greater than the activities measured in corresponding methanol and acetonitrile aqueous solutions. This biophysical study shows that AAFs have potential application as organic solvent replacements at moderate temperature in the mobile phase for the separation of proteins in their native form by reversed phase liquid chromatography.
登录
查看更多内容
影响因子：
2.9
作者：
GOTO, Y;HAGIHARA, Y;NISHII, I
通讯作者：
NISHII, I
DOI：
10.1016/j.chroma.2008.09.064
发表时间：
2009-04-17
期刊：
Journal of chromatography. A
影响因子：
--
作者：
Grossman S;Danielson ND
通讯作者：
Danielson ND
影响因子：
2.9
作者：
Huang, Mian-Mian;Weingaertner, Hermann
通讯作者：
Weingaertner, Hermann
影响因子：
5.6
作者：
Byrne, N.;Angell, C. A.
通讯作者：
Angell, C. A.
影响因子：
15
作者：
Lyubovitsky, JG;Gray, HB;Winkler, JR
通讯作者：
Winkler, JR