CAJ | 학술논문

目的探讨血小板膜糖蛋白(GP)Ⅰ bα胞内区551到565氨基酸序列对GPⅠ bα结合血管性血友病因子(VWF)功能的调控作用。方法以同时表达野生型GPⅠ bα、GP Ⅰ bβ和GPⅨ三种蛋白的中国仓鼠卵巢细胞株(1b9)、同时表达野生型GP Ⅰ bβ、GPⅨ和在565或551以后氨基酸序列缺失的GPⅠ bα的中国仓鼠卵巢细胞株(△565或△551)为模型，采用流式细胞术检测细胞株的GP Ⅰbα在瑞斯托霉素诱导下结合VWF的能力，流动腔技术检测细胞株在流体剪切力条件下(200 s-1)在VWF表面的黏附状况，激光共聚焦技术检测细胞株在botrocetin诱导下在VWF表面的铺展状况。结果与对照1b9和△551相比，△565细胞株在ristocetin诱导下其GPⅠ bα结合VWF的能力最强(P＜0.01)，在VWF表面黏附的△565细胞数量最多(P＜0.05)，在botrocetin诱导下△565细胞株在VWF表面的铺展面积最大(P＜0.05)。结论 GP Ⅰ bα胞内区551到565氨基酸序列对GP Ⅰ bα结合VWF的功能具有重要调控作用。
목적 탐토혈소판막당단백(GP)Ⅰ bα포내구551도565안기산서렬대GPⅠ bα결합혈관성혈우병인자(VWF)공능적조공작용。방법 이동시표체야생형GPⅠ bα、GP Ⅰ bβ화GPⅨ삼충단백적중국창서란소세포주(1b9)、동시표체야생형GP Ⅰ bβ、GPⅨ화재565혹551이후안기산서렬결실적GPⅠ bα적중국창서란소세포주(△565혹△551)위모형，채용류식세포술검측세포주적GP Ⅰbα재서사탁매소유도하결합VWF적능력，류동강기술검측세포주재류체전절력조건하(200 s-1)재VWF표면적점부상황，격광공취초기술검측세포주재botrocetin유도하재VWF표면적포전상황。결과 여대조1b9화△551상비，△565세포주재ristocetin유도하기GPⅠ bα결합VWF적능력최강(P＜0.01)，재VWF표면점부적△565세포수량최다(P＜0.05)，재botrocetin유도하△565세포주재VWF표면적포전면적최대(P＜0.05)。결론 GP Ⅰ bα포내구551도565안기산서렬대GP Ⅰ bα결합VWF적공능구유중요조공작용。
Objective To explore the role of the amino acids between 551 and 565 in the cytoplasmic domain of glycoprotein (GP) Ⅰ bα in the VWF binding to GP Ⅰ bα. Methods The VWF binding to GP Ⅰ bα induced by ristocetin was analyzed by flow cytometry, in three GP Ⅰ b-Ⅸ -expressing Chinese hamster ovary (CHO) cell lines 1b9, △565 and △551, adhesion of above cells on VWF by flow chamber analysis at shear rate of 200 s-1. The spread of GP Ⅰ b-Ⅸ -expressing cells were stimulated with botrocetin on VWF-coated coverslips by confocal microscope. Results The VWF binding to GP Ⅰ bα was higher in △565 cells stimulated by ristocetin than in △551 or 1b9 cells. The number of △565 cells adhered on the VWF-coatedchamber was more than that of controls at shear rate of 200 s-1. Moreover, the surface spreading areas of △565 cells were greater than that of the controls on VWF-coated coverslips. Conclusions The amino acids between 551 and 565 in the cytoplasmic domain of GP Ⅰ bα regulates the VWF binding to GP Ⅰ bα.