Carter R H, Tuveson D A, Park D J, Rhee S G, Fearon D T
Division of Molecular and Clinical Rheumatology, Johns Hopkins University School of Medicine, Baltimore, MD 21205.
J Immunol. 1991 Dec 1;147(11):3663-71.
We have investigated the mechanism by which the membrane protein complex of the B lymphocyte that contains CD19 and CR2 activates phospholipase C (PLC) to induce a rise in [CA2+]i. The CD19 complex resembled the membrane IgM complex in that three protein tyrosine kinase inhibitors suppressed increases in [Ca2+]i and inositol bisphosphate and inositol triphosphate generation. However, the activation of PLC by the CD19 complex could be distinguished from that by the membrane IgM complex by slower kinetics of generation of inositol phosphates, resistance to inhibition by activators of protein kinase C, and different pattern of tyrosine-phosphorylated cellular substrates. Western blot analysis of lysates from cells stimulated by the CD19 complex demonstrated a single new phosphotyrosine-containing protein of 85 kDa, whereas multiple other phosphotyrosine-containing proteins were present in cells activated by the mIgM complex. In particular, PLC-gamma 1, which is a substrate for the protein tyrosine kinase activated by the mIgM complex, was not tyrosine-phosphorylated in cells stimulated by the CD19 complex. Cross-linking the two complexes together caused a synergistic increase in [CA2+]i which was neither suppressed by activation of protein kinase C nor associated with increased tyrosine-phosphorylation of PLC, characteristic of the CD19 pathway. Therefore, the B cell has two signal transduction complexes, associated with membrane IgM and CD19, that activate PLC by different mechanisms and that can synergistically interact to enhance this function by the CD19 pathway.
我们研究了含有CD19和CR2的B淋巴细胞膜蛋白复合物激活磷脂酶C(PLC)以诱导细胞内钙离子浓度([Ca2+]i)升高的机制。CD19复合物与膜IgM复合物相似之处在于,三种蛋白酪氨酸激酶抑制剂可抑制[Ca2+]i的升高以及肌醇二磷酸和肌醇三磷酸的生成。然而,CD19复合物对PLC的激活与膜IgM复合物对PLC的激活可通过以下方面加以区分:肌醇磷酸生成的动力学较慢、对蛋白激酶C激活剂的抑制具有抗性以及酪氨酸磷酸化细胞底物的模式不同。对受CD19复合物刺激的细胞裂解物进行的蛋白质印迹分析显示,有一种新的85 kDa含磷酸酪氨酸的单一蛋白,而在由mIgM复合物激活的细胞中存在多种其他含磷酸酪氨酸的蛋白。特别是,作为被mIgM复合物激活的蛋白酪氨酸激酶底物的PLC-γ1,在受CD19复合物刺激的细胞中未发生酪氨酸磷酸化。将这两种复合物交联在一起会导致[Ca2+]i协同增加,这种增加既不被蛋白激酶C的激活所抑制,也与PLC酪氨酸磷酸化增加无关,这是CD19途径的特征。因此,B细胞有两种与膜IgM和CD19相关的信号转导复合物,它们通过不同机制激活PLC,并且可以通过CD19途径协同相互作用以增强该功能。