Milligan G
Department of Biochemistry, University of Glasgow, U.K.
Biochem J. 1987 Jul 15;245(2):501-5. doi: 10.1042/bj2450501.
Cellular proliferation of rat glioma C6 BU1 cells in tissue culture is dependent on the presence of either calf or foetal-calf serum in the medium. Foetal-calf serum stimulated a high-affinity GTPase in membranes derived from C6 BU1 cells. Pretreatment of the cells with pertussis toxin decreased the high-affinity GTPase activity substantially, and attenuated the foetal-calf-serum-stimulated increase in this GTPase activity. Cholera toxin, in contrast, did not modulate the response to foetal-calf serum. Foetal-calf serum did not inhibit adenylate cyclase activity in membranes of these cells, indicating that the G-protein that was stimulated by foetal-calf serum was not Gi (the inhibitory one). Although the nature of the specific component of foetal-calf serum responsible for this pertussis-toxin-sensitive receptor-mediated stimulation of high-affinity GTPase activity has not been identified, it was mimicked neither by bombesin, which can stimulate inositol phospholipid turnover via a guanine nucleotide binding protein, nor by platelet-derived growth factor, which is present in substantial concentrations in foetal-calf serum. This report represents the first demonstration of a pertussis-toxin-substrate-mediated response in this cell line and provides further evidence that G-proteins other than Gi can be functionally inactivated by pertussis toxin.
大鼠胶质瘤C6 BU1细胞在组织培养中的细胞增殖依赖于培养基中存在小牛血清或胎牛血清。胎牛血清刺激了C6 BU1细胞来源膜中的一种高亲和力GTP酶。用百日咳毒素预处理细胞可显著降低高亲和力GTP酶活性,并减弱胎牛血清刺激的该GTP酶活性增加。相比之下,霍乱毒素并未调节对胎牛血清的反应。胎牛血清并未抑制这些细胞的膜中的腺苷酸环化酶活性,这表明受胎牛血清刺激的G蛋白不是Gi(抑制性G蛋白)。尽管尚未确定胎牛血清中负责这种百日咳毒素敏感的受体介导的高亲和力GTP酶活性刺激的特定成分的性质,但蛙皮素(其可通过鸟嘌呤核苷酸结合蛋白刺激肌醇磷脂周转)和血小板衍生生长因子(其在胎牛血清中大量存在)均未模拟该作用。本报告首次证明了该细胞系中存在百日咳毒素底物介导的反应,并进一步证明除Gi外的其他G蛋白可被百日咳毒素功能性失活。