6A8 cDNA编码一种α-甘露糖苷酶

6A8 cDNA Encodes A Type of α-mannosidase

目的 探讨6A8cDNA编码的蛋白质的性质。方法 依据ConA的配体是甘露糖,而α-甘露糖苷酶的作用是修剪细胞糖蛋白聚糖中甘露糖的原理,采用细胞免疫荧光染色法以及激光共聚焦显微镜观察经转染6A8cDNA正义或反义重组表达载体后,细胞结合ConA的强弱,验证6A8cDNA编码的蛋白质是否为一种α-甘露糖苷酶。结果 COS-7细胞和人鼻咽癌细胞株CNE-2L2转染pRc/CMV-正义6A8cDNA后与ConA结合减弱,而与单抗6A8染色增强;CNE-2L2细胞转染pRc/CMV-反义6A8cDNA后,与ConA结合增强,而与单抗6A8染色减弱。结论 6A8cDNA编码的蛋白质可能为一种α-甘露糖苷酶,它可被单抗6A8识别。

To find the nature of the protein encoded by 6A8 cDNA. Methods In the light of that the ligand of Con A is mannoses and that the function of α mannosidase is trimming of mannoses in the oligosaccharide chains of glycoproteins, COS 7 cells and CNE 2L2 cells, a cloned cell line of human nasopharyngeal carcinoma, were transfected with pRc/CMV sense 6A8 cDNA or pRc/CMV antisense 6A8 cDNA and the Con A binding to the cells was examined upon a lazer scan confocal microscope. Results The data demonstrated that with transfection of pRc/CMV sense 6A8 cDNA the binding of Con A to the cells was decreased, but the staining of the cells with mAb 6A8 was enhanced, and vise versa for the cells with transfection of pRc/CMV antisense 6A8 cDNA. Conclusions The protein encoded by 6A8 cDNA might be a type of α mannosidase recognized by mAb 6A8.