Label-free chiral detection of melittin binding to a membrane
Label-free chiral detection of melittin binding to a membrane
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DOI：
10.1021/ja028577z
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发表时间：
2003-02-05
影响因子：
15
通讯作者：
Conboy, JC
中科院分区：
文献类型：
--
作者：
Kriech, MA;Conboy, JC
本研究描述了一种利用手性二次谐波产生（C-SHG）检测表面受限蛋白的创新方法。采用了一种独特的光学几何结构，可以分离手性和非手性非线性响应。利用这种光学排列，C-SHG首次检测了蜂毒素吸附在平面支撑的脂质双分子层上的手性。通过检测C-SHG信号，监测蜂毒素与膜的结合作为体积浓度的函数。C-SHG吸附等温线分析表明，其吸附行为具有正相互作用能。测定得到的结合常数Ka为（8.3±1.0）× 105M-1。这些研究结果对利用C-SHG进行蛋白与表面结合的无标记检测和蛋白界面现象的分析具有深远的意义。
The study presented here describes an innovative approach for the detection of surface-confined proteins using chiral second harmonic generation (C-SHG). A unique optical geometry has been employed which allows for the separation of the chiral and achiral nonlinear response. By utilizing this optical arrangement, the detection of chirality originating from melittin adsorbed to a planar supported lipid bilayer has been performed for the first time by C-SHG. Melittin binding to the membrane was monitored as a function of bulk concentration through detection of the C-SHG signal. Analysis of the C-SHG adsorption isotherms reveals Frumkin adsorption behavior with a positive interaction energy. The binding constant (Ka) obtained was determined to be (8.3 ± 1.0) × 105M-1. The results of these studies have far-reaching implication in the use of C-SHG for the label-free detection of protein association to surfaces and in the analysis of protein interfacial phenomena.