Binding of doxyl stearic spin labels to human serum albumin: an EPR study.

Binding of doxyl stearic spin labels to human serum albumin: an EPR study.
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硬脂酸自旋标签与人血清白蛋白的结合:一项 EPR 研究。

DOI:
10.1021/jp5068928
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发表时间:
2014
期刊:
The journal of physical chemistry. B
影响因子:
--
通讯作者:
G. Bačić
G. Bačić
中科院分区:
--
文献类型:
--
作者:
A. Pavićević;A. Popović;M. Mojović;Snežana V Šušnjar;G. Bačić

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通过电子顺磁共振 (EPR) 光谱检查自旋标记脂肪酸 (SLFA) 与人血清白蛋白 (HSA) 的结合,以评估 HSA/SLFA/EPR 技术作为癌症生物标记工具的潜力。对脂肪酸 (FA) 链两端连接有硝基氧基团的两种自旋标签 5-doxyl 硬脂酸 (5-DS) 和 16-doxyl 硬脂酸 (16-DS) 进行了比较研究。孵育时间、不同[SLFA]/[HSA]摩尔比、乙醇和温度的影响表明,硝基氧基团的位置使两种SLFA之间的结合产生一定的差异。不同[SLFA]/[HSA]摩尔比的光谱被分解为两个光谱分量,分别对应于弱结合和强结合的SLFA。抗坏血酸对 SLFA 的还原表明存在快速和慢速的双组分过程,证实了分解结果。华法林对两种 SLFA 的结合没有影响,而布洛芬显着降低 5-DS 的结合,对 16-DS 没有影响。总之,这项研究的结果表明,SLFA、5-DS 和 16-DS 都应该用于研究各种医疗条件引起的血液中 HSA 构象变化。
The binding of spin-labeled fatty acids (SLFAs) to the human serum albumin (HSA) examined by electron paramagnetic resonance (EPR) spectroscopy was studied to evaluate the potential of the HSA/SLFA/EPR technique as a biomarking tool for cancer. A comparative study was performed on two spin labels with nitroxide groups attached at opposite ends of the fatty acid (FA) chain, 5-doxyl stearic (5-DS) and 16-doxyl stearic (16-DS) acid. The effects of incubation time, different [SLFA]/[HSA] molar ratios, ethanol, and temperature showed that the position of the nitroxide group produces certain differences in binding between the two SLFAs. Spectra for different [SLFA]/[HSA] molar ratios were decomposed into two spectral components, which correspond to the weakly and strongly bound SLFAs. The reduction of SLFA with ascorbate showed the existence of a two component process, fast and slow, confirming the decomposition results. Warfarin has no effect on the binding of the two SLFAs, whereas ibuprofen significantly decreases the binding of 5-DS and has no effect on 16-DS. Together, the results of this study indicate that both SLFAs, 5-DS and 16-DS, should be used for the study of HSA conformational changes in blood induced by various medical conditions.
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