Exploring the pH-Induced Functional Phase Space of Human Serum Albumin by EPR Spectroscopy
Exploring the pH-Induced Functional Phase Space of Human Serum Albumin by EPR Spectroscopy
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DOI：
10.3390/magnetochemistry4040047
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发表时间：
2018-12-01
期刊：
影响因子：
2.7
通讯作者：
Hinderberger, Dariush
中科院分区：
文献类型：
--
作者：
Reichenwallner, Joerg;Oehmichen, Marie-T.;Hinderberger, Dariush
用电子顺磁共振（EPR）方法研究了人血清白蛋白（HSA）与顺磁性羟基硬脂酸（5-DSA和16-DSA）配体的自组装溶液体系。它是测试在何种程度上的pH值诱导的构象异构体的HSA显示自己在连续波（CW）EPR光谱从这种自旋探测方法相比，一个既定的自旋标记策略，利用3-马来酰亚胺基（5-MSL）。大多数分析都是在经验水平上进行的，具有强大的策略，可以检测配体和蛋白质的动态变化。特别强调的是已被放置在EPR光谱检测的熔融球（MG）状态的HSA，通常发现的荧光探针8-苯胺基-萘-1-磺酸（ANS）。此外，四脉冲双电子-电子共振（DEER）实验进行和证实与动态光散射（DLS）的数据，以确定在溶液形状的HSA与pH值的变化。所有的结果最终结合在一个详细的计划，描述了pH值引起的功能相空间的HSA。
A systematic study on the self-assembled solution system of human serum albumin (HSA) and paramagnetic doxyl stearic acid (5-DSA and 16-DSA) ligands is reported covering the broad pH range 0.7-12.9, mainly using electron paramagnetic resonance (EPR) methods. It is tested to which extent the pH-induced conformational isomers of HSA reveal themselves in continuous wave (CW) EPR spectra from this spin probing approach in comparison to an established spin-labeling strategy utilizing 3-maleimido proxyl (5-MSL). Most analyses are conducted on empirical levels with robust strategies that allow for the detection of dynamic changes of ligand, as well as protein. Special emphasis has been placed on the EPR spectroscopic detection of a molten globule (MG) state of HSA that is typically found by the fluorescent probe 8-Anilino-naphthalene-1-sulfonic acid (ANS). Moreover, four-pulse double electron-electron resonance (DEER) experiments are conducted and substantiated with dynamic light scattering (DLS) data to determine changes in the solution shape of HSA with pH. All results are ultimately combined in a detailed scheme that describes the pH-induced functional phase space of HSA.