Sármay G, Koncz G, Gergely J
Laboratory of Immunoregulation, Vienna International Research Cooperation Center at SFI, Vienna, Austria.
J Biol Chem. 1996 Nov 29;271(48):30499-504. doi: 10.1074/jbc.271.48.30499.
Co-ligation of antigen receptors and type II Fcgamma receptors (FcgammaRIIb) on B cells interrupts signal transduction and ultimately inhibits antibody production. We have identified p52 Shc in the FcgammaRIIb1-specific immunoprecipitates isolated from the membrane fraction of BL41 Burkitt lymphoma cells following B cell receptor-FcgammaRIIb1 co-ligation. The insolubilized synthetic peptide representing the phosphorylated form of the tyrosine-based inhibitory motif of FcgammaRIIb also binds Shc from the lysates of activated but not from resting BL41 cells. This suggests that the binding does not depend on the interaction of FcgammaRIIb1-phosphotyrosine with the SH2 domain of Shc. Tyr phosphorylation of FcgammaRIIb1-associated Shc is low, indicating an impaired function. Shc is implicated in regulating p21(ras) activation; thus, we have compared p21(ras) activities in BL41 cells treated in different ways. p21(ras) activity is reduced when B cell receptor and FcgammaRIIb1 are co-ligated. p21(ras) couples protein-tyrosine kinase-dependent events to the Ser/Thr kinase-mediated signaling pathway leading to the activation of mitogen-activated protein kinases (MAPK). Our results show that B cell receptor-FcgammaRIIb1 co-cross-linking partially inhibits mitogen-activated protein kinase activity. We conclude that FcgammaRIIb1-dependent inhibition of human B cell activation may be based on interrupting signal transduction between protein-tyrosine kinases and the p21(ras)/mitogen-activated protein kinase-dependent activation pathway.
B细胞上抗原受体与II型Fcγ受体(FcγRIIb)的共连接会中断信号转导,并最终抑制抗体产生。我们在B细胞受体 - FcγRIIb1共连接后,从BL41伯基特淋巴瘤细胞膜组分中分离出的FcγRIIb1特异性免疫沉淀物中鉴定出了p52 Shc。代表FcγRIIb基于酪氨酸的抑制基序磷酸化形式的不溶性合成肽也能从活化的BL41细胞裂解物中结合Shc,但不能从静止的BL41细胞裂解物中结合。这表明这种结合不依赖于FcγRIIb1 - 磷酸酪氨酸与Shc的SH2结构域之间的相互作用。与FcγRIIb1相关的Shc的酪氨酸磷酸化水平较低,表明其功能受损。Shc参与调节p21(ras)的激活;因此,我们比较了以不同方式处理的BL41细胞中的p21(ras)活性。当B细胞受体和FcγRIIb1共连接时,p21(ras)活性降低。p21(ras)将蛋白酪氨酸激酶依赖性事件与丝氨酸/苏氨酸激酶介导的信号通路偶联,导致丝裂原活化蛋白激酶(MAPK)的激活。我们的结果表明,B细胞受体 - FcγRIIb1共交联部分抑制丝裂原活化蛋白激酶活性。我们得出结论,FcγRIIb1依赖性对人B细胞激活的抑制可能基于中断蛋白酪氨酸激酶与p21(ras)/丝裂原活化蛋白激酶依赖性激活途径之间的信号转导。