使用特异性单克隆抗体研究整合膜蛋白从分泌颗粒经分泌型大鼠嗜碱性白血病细胞膜的转运过程。
Study of the transit of an integral membrane protein from secretory granules through the plasma membrane of secreting rat basophilic leukemia cells using a specific monoclonal antibody.
作者信息
Bonifacino J S, Perez P, Klausner R D, Sandoval I V
出版信息
J Cell Biol. 1986 Feb;102(2):516-22. doi: 10.1083/jcb.102.2.516.
Abstract
The monoclonal antibody 5G10 reacted specifically with an 80-kD integral membrane protein in rat basophilic leukemia (RBL) cells. Immunofluorescence microscopy studies of RBL cells, fixed and permeabilized, revealed that the 80-kD protein was located in the membrane of cytoplasmic vesicles. The vesicles were identified as secretory granules by their content in immunoreactive serotonin. Expression of the 5G10 antigen on the surface of unstimulated RBL cells was low. However, RBL cells stimulated to secrete with anti-dinitrophenyl IgE followed by dinitrophenyl-bovine serum albumin or with the Ca2+ ionophore A-23187 displayed an increased expression of the antigen on their surface. Surface exposure of the 5G10 antigen was maximal at 5 min after stimulation of secretion. Removal of dinitrophenyl-bovine serum albumin from the incubation medium resulted in internalization of 50% of the antigen within 10 min.
摘要
单克隆抗体5G10与大鼠嗜碱性白血病（RBL）细胞中的一种80-kD整合膜蛋白发生特异性反应。对固定并通透处理的RBL细胞进行免疫荧光显微镜研究发现，80-kD蛋白位于细胞质囊泡膜上。这些囊泡因其含有免疫反应性血清素而被鉴定为分泌颗粒。未受刺激的RBL细胞表面5G10抗原的表达较低。然而，用抗二硝基苯基IgE刺激RBL细胞分泌，随后加入二硝基苯基牛血清白蛋白，或用Ca2+离子载体A-23187刺激，其表面抗原表达增加。分泌刺激后5分钟，5G10抗原的表面暴露达到最大值。从孵育培养基中去除二硝基苯基牛血清白蛋白会导致10分钟内50%的抗原内化。
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