Analysis of factor VIIa binding to relipidated tissue factor by surface plasmon resonance.
Analysis of factor VIIa binding to relipidated tissue factor by surface plasmon resonance.
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DOI：
10.1097/mbc.0b013e328333b084
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发表时间：
2010-06
期刊：
影响因子：
--
通讯作者：
Rao LV
中科院分区：
文献类型：
--
作者：
Sen P;Neuenschwander PF;Pendurthi UR;Rao LV
组织因子(Tf)-FVIIa(FVIIa)相互作用的动力学分析有助于研究Tf-FVIIa的结构-功能关系。然而，已报道的FVIIa与Tf的结合亲和力之间存在着很大的差异，特别是当比较功能活性分析和配体结合研究获得的Kd值时。表面等离子体共振(SPR)技术常被用来研究无脂环境中FVIIa与Tf的结合动力学。在目前的研究中，我们使用包埋在磷脂双层中的转铁蛋白，通过SPR来确定结合运动。数据表明，FVIIa与磷脂双层中的Tf的结合亲和力比无脂环境中的Tf高出100倍，接近酶活性测定中的Kd值。目前的数据表明，利用磷脂中嵌入的Tf进行SPR结合研究更适合于研究FVIIa(或FVIIa突变体/衍生物)在生理环境中如何与Tf相互作用。
Kinetic analysis of the tissue factor (TF)-factor VIIa (FVIIa) binding interaction is helpful in investigating the structure-function relationships of TF-FVIIa. However, a wide variation exists among the reported binding affinities of FVIIa to TF, particularly when comparing KD values obtained from functional activity assays versus ligand binding studies. Surface plasmon resonance (SPR) technique was used frequently to investigate binding kinetics of FVIIa to TF in a lipid-free environment. In the present study we used TF embedded in a phospholipid bilayer for determining binding kinectis using SPR. The data revealed that FVIIa had a much higher binding affinity (>100-fold) for TF embedded in the phospholiid bilayer than TF in a lipid-free environment, approaching the KD values that were noted in the enzymatic activity assays. The present data suggest that SPR binding studies using TF embedded in phopsholipids is more appropritate for investigating how FVIIa (or FVIIa mutants/derivatives) may interact with TF in physiological settings.