亲和毛细管电泳法研究锌离子与人血清白蛋白的结合反应机制

建立了研究金属离子与人血清白蛋白(Human serum albumin,HSA)相互作用的亲和毛细管电泳(Affinity capillary electrophoresis,ACE)方法。生理条件下,构建配体(Zn2+)-受体(HSA)相互作用模型,以N,N-二甲基甲酰胺(N,N-Dimethylformamide,DMF)为内标物,基于Scatchard方程,依据有效淌度的变化,通过非线性模拟方程计算Zn2+-HSA结合反应的表观结合常数KB,定量表征了Zn2+-HSA相互作用的强度,并解析电泳谱图获得了Zn2+-HSA结合反应为一快平衡体系的结论。结果表明,建立的ACE方法简捷、有效,Zn2+-HSA相互作用的强度与Zn2+浓度之间存在明显的量效关系。

Binding Interaction Between Zinc Ion and Human Serum Albumin Using Affinity Capillary Electrophoresis

The interaction between metal ions and human serum albumin(HSA) was investigated using affinity capillary electrophoresis(ACE) method.A model about the interaction between zinc ion as the ligand and HSA as the receptor was established under physiological conditions.The changes of effective mobility were determined for this interaction system with N,N-dimethylformamide(DMF) as internal marker.Based on the Scatchard model,a practical nonlinear simulation equation was used to calculate the apparent binding constant and the interaction binding strength between metal ion and HSA was quantificational characterized,and a conclusion about a quick balance system of the interaction between metal ion and HSA was acquired by analyzing electropherogram.It was concluded that ACE method was simple and effective and the obvious response relationship existed between the interaction strength and the concentrations of zinc ion.This work has referential meaning for understanding deeply the interaction strength between metal ion and HSA in vivo.The affinity capillary electrophoresis method can be considered valid for studying the interaction between metal ions and protein molecules.