School of Life Science, Beijing Institute of Technology, Beijing, P. R. China.
Electrophoresis. 2011 Mar;32(6-7):749-51. doi: 10.1002/elps.201000618. Epub 2011 Mar 1.
This short communication describes the interaction between toll-like receptor 4 (TLR4) and lipopolysaccharide (LPS, its specific ligand) using analytical methods. Their interaction has been evidenced in many reports. Nevertheless, there are few reports focused on their binding constant. In this research, the interaction between TLR4 and LPS is investigated using mobility shift method by CZE. To optimize the electrophoresis conditions, the effecting factors, running buffer, sample concentration, injection duration, and operation voltage of electrophoretic on the mobility shift are studied in detail. Electrophoresis conditions were described as follows: borate buffer (pH 7.4, 20 mM), 5 s for 50 mbar pressure injection duration, and 13 kV of separation voltage in 41.5 cm fused silica capillary with 75 μm id and 375 μm od. The combination constant of TLR4 and LPS is calculated using Scatchard methods. The Scatchard liner correlation is y=-0.0165x+0.1456, binding constant is K=1.65 x 10⁴ (g/mL)⁻¹.
本短文通讯使用分析方法描述了 Toll 样受体 4（TLR4）与脂多糖（LPS，其特定配体）之间的相互作用。它们之间的相互作用在许多报道中都有证据。然而，很少有报道关注它们的结合常数。在这项研究中，使用 CZE 的迁移率变化方法研究了 TLR4 和 LPS 之间的相互作用。为了优化电泳条件，详细研究了电泳条件对迁移率变化的影响因素，包括运行缓冲液、样品浓度、进样时间、操作电压。电泳条件描述如下：硼酸盐缓冲液（pH 7.4，20 mM），50 mbar 压力进样时间为 5 s，在 41.5 cm 熔融硅毛细管中以 75 μm id 和 375 μm od 进行 13 kV 的分离电压。使用 Scatchard 方法计算 TLR4 和 LPS 的结合常数。Scatchard 线性相关为 y=-0.0165x+0.1456，结合常数为 K=1.65 x 10⁴ （g/mL）⁻¹。
