A surface plasmon resonance assay for the binding of influenza virus hemagglutinin to its sialic acid receptor.
A surface plasmon resonance assay for the binding of influenza virus hemagglutinin to its sialic acid receptor.
复制标题
DOI：
10.1006/viro.1996.0139
复制
发表时间：
1996-03
期刊：
影响因子：
3.7
通讯作者：
Darin K. Takemoto;J. Skehel;D. Wiley;D. Wiley
中科院分区：
文献类型：
--
作者：
Darin K. Takemoto;J. Skehel;D. Wiley;D. Wiley
我们开发了一种灵敏的微尺度结合试验，利用实时表面等离子体共振来研究流感血凝素与其细胞表面受体唾液酸之间的相互作用。用n -羟基琥珀酰亚胺和n -乙基- n '-（二甲氨基丙基）碳二亚胺将糖蛋白fetuin结合到羧甲基化的葡聚糖传感器表面。低ph诱导的BHA莲座特异性结合胎儿素衍生的传感器表面，但不结合asialofetun衍生的传感器表面。结合可以通过用毫摩尔浓度的流感血凝素-唾液酸相互作用抑制剂预先孵育BHA莲座来抑制。我们还测量了BHA莲座与胎蛋白衍生传感器表面之间多价相互作用的结合率、解离率和解离常数，从而量化了通过BHA莲座与胎蛋白衍生传感器表面之间的多价相互作用实现的紧密结合。
We have developed a sensitive microscale binding assay to study the interaction between influenza hemagglutinin and its cell surface receptor sialic acid using real-time surface plasmon resonance. The glycoprotein fetuin was bound to a carboxymethylated-Dextran sensor surface using N-hydroxysuccinimide and N-ethyl-N'-(dimethylaminopropyl) carbodiimide. Low-pH-induced BHA rosettes bind specifically to the fetuin-derivitized sensor surface, but not to an asialofetuin-derivitized sensor surface. Binding can be inhibited by preincubation of BHA rosettes with millimolar concentrations of inhibitors of the influenza hemagglutinin-sialic acid interaction. The association rate, dissociation rate, and dissociation constant for the multivalent interaction between BHA rosettes and the fetuin-derivitized sensor surface were also measured, allowing us to quantitate the tight binding achieved through the multivalent interaction between BHA rosettes and the fetuin-derivitized sensor surface.