Identification of the peptides that stimulate the phosphoinositide hydrolysis in lymphocyte cell lines from peptide libraries
Identification of the peptides that stimulate the phosphoinositide hydrolysis in lymphocyte cell lines from peptide libraries
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DOI：
10.1074/jbc.271.14.8170
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发表时间：
1996-04-05
影响因子：
4.8
通讯作者：
Ryu, SH
中科院分区：
文献类型：
--
作者：
Baek, SH;Seo, JK;Ryu, SH
通过筛选由随机序列的六肽组成的合成肽文库，鉴定了刺激淋巴细胞系中肌醇磷酸(INOPs)形成的多肽。含有相同序列XKYX(P/V)M的多肽在磷脂酶C(PLC)介导的人B骨髓瘤细胞株U266中iNOPs的形成中活性最高，在HL60和U937细胞中也能刺激肌醇磷脂的水解和[Ca+](I)的释放，而对以下细胞株NIH3T3、PC12、Daudi、Sp2、Jurkat、H9、Molt-4、Supt-1、K562和RBL-2H3则没有影响。这一结果提示，这些多肽可能具有细胞类型特异性。其中一种活性多肽WKYMVM-NH2的作用与G蛋白激活剂AlF4的作用相似，百日咳毒素部分阻断了其诱导的iNOPs的聚集和[Ca~(2+)]的释放。与I-125标记的多肽的结合分析表明，U266细胞具有相当数量的多肽结合位点。这些结果表明，在某些细胞类型中，百日咳毒素敏感的G-蛋白偶联受体可激活PLC介导的信号转导。
Peptides which stimulate the formation of inositol phosphates (InoPs) in lymphocyte cell lines were identified by screening synthetic peptide libraries composed of random sequences of hexapeptides. The peptides containing the consensus sequence XKYX(P/V)M were found to be most active in the phospholipase C (PLC)-mediated formation of InoPs in a human B myeloma cell line, U266, The peptides also stimulated the phosphoinositide hydrolysis and the release of [Ca2+](i) in HL60 and U937 cell lines, On the other hand, these peptides showed no effect in the following cell lines: NIH3T3, PC12, Daudi, Sp2, Jurkat, H9, Molt-4, SupT-1, K562, and RBL-2H3. The result suggests the possibility that the peptides may have cell type specificity. Experiments with one of the active peptides, WKYMVM-NH2 showed that its action mimics the effect of AlF4- which is a G-protein activator in the InoPs generation, and pertussis toxin partially blocked the InoPs accumulation and [Ca2+](i) release induced by the peptide in the U266 cells, Binding assays with the peptide labeled with I-125 showed that U266 cells have a saturable number of binding sites for the peptide, Taken together, these results suggest that the peptides could activate PLC-mediated signal transduction via a pertussis toxin-sensitive G-protein coupled receptor in certain cell types.