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本实验用微吸管技术研究血管性血友病因子(von Willebrand factor,VWF)与血小板膜糖蛋白(GPIb/IX)之间相互识别、黏附的反应率及两种蛋白黏附之后发生解离时所需要的解离力。利用经过转染可表达人血小板膜糖蛋白GPIb/IX的CHO细胞与包被有VWF的兔红细胞互相接触并施加一定的外界压力,保持压力一段时间之后根据分开两细胞时红细胞变形判断蛋白质识别与否,记录两种蛋白质发生识别黏附的几率。通过对细胞变形的测量和数据处理得出GPIb/IX与VWF识别后的解离力。携带GPIb/IX或VWF的细胞对心挤压0.5μm,持续30s的实验状态下,两个细胞间发生黏附的概率约为15.0%。通过对红细胞形变的分析,得到携带GPIb/IX或VWF的细胞发生黏附后的解离力呈随机分布状态。分布的峰值,即解离力的最大几率值为10 pN。
In this study, the micro-pipette technique was used to study the relationship between von Willebrand factor (VWF) and platelet glycoprotein (GPIb / IX), the reaction rate of adhesion and the dissociation of two proteins after their adhesion The required force of dissociation. CHO cells transfected with GPIb / IX expressing human platelet membrane glycoprotein and rabbit erythrocytes coated with VWF were contacted with each other and subjected to a certain external pressure, and the pressure was maintained for a period of time to judge the protein identification based on the erythrocyte deformation when the two cells were separated No, record the probability that two proteins will recognize adhesion. Through the measurement of cell deformation and data processing GPIb / IX and VWF identified dissociation force. Cells carrying GPIb / IX or VWF had a cardiac arrest of 0.5 μm for an experimental condition of 30 s, with a probability of about 15.0% adhesion between the two cells. Through the analysis of erythrocyte deformability, we found that the dissociation forces of cells carrying GPIb / IX or VWF were randomly distributed. The peak value of the distribution, the maximum probability of dissociation, is 10 pN.