Study of the role of ectokinase on the signal transduction in mast cells

外激酶对肥大细胞信号转导作用的研究

• 批准号: 09672270
• 负责人: TESHIMA Reiko
• 金额: $ 1.79万
• 依托单位: National Institute of Health Sciences
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 1997
• 资助国家: 日本
• 起止时间: 1997 至 1998
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-09672270/
• 关键词: IgE receptor; Ectokinases; Mast cells; Signal transduction; Degranulation; Calcium signal

We examined the effects of K252b, an ectoprotein kinase inhibitor of microbial origin, on the activation of RBL-2H3 cells by cross-linking of IgB receptors or by the Ca^<2+> ionophore A23187. Analysis of phosphorylation of ectoproteins following IgE receptor cross-linking revealed that K252b mainly inhibited the phosphorylation of a 130-kDa protein. The inhibitor simultaneously inhibited the degranulation and the sustained increase in the cytosolic calcium ion concentration even afteraddition of Ag. In contrast, K252b did not inhibit the increase in degranulation and cytosolic calcium ion concentration caused by stimulation with A23187. Permeation of K252b into RBL-2H3 cells, assessed by fluorescence intensity, was very low. K252b also inhibited de-granulation caused by IgE receptor cross-linking in human basophils, but did not inhibit the de-granuation caused by A23187. Thus, our findings suggest that the effects of K252b may be mediated by outersurface-bound or -anchored K252b-sensitive molecules on RBL-2H3 cells and human basophils, and that the phosphorylation of ectoprotein may be involve a transmembrane influx of Ca^<2+> by IgE rece-ptor cross-linking.

我们研究了微生物来源的外蛋白激酶抑制剂K252b通过IgB受体交联或Ca^<2+>离子载体A23187对RBL-2H3细胞的激活作用。对IgE受体交联后外蛋白磷酸化的分析显示，K252b主要抑制130-kDa蛋白的磷酸化。该抑制剂同时抑制了脱粒，并在添加银后持续增加胞质钙离子浓度。相比之下，K252b不抑制A23187刺激引起的脱颗粒和胞质钙离子浓度的增加。通过荧光强度测定，K252b对RBL-2H3细胞的渗透非常低。K252b对人嗜碱性细胞中IgE受体交联引起的脱粒也有抑制作用，但对A23187引起的脱粒没有抑制作用。因此，我们的研究结果表明，K252b的作用可能是由外表面结合或锚定的K252b敏感分子介导的，这些分子对rbr - 2h3细胞和人嗜碱性细胞起作用，并且外显蛋白的磷酸化可能涉及IgE受体交联介导Ca^<2+>的跨膜内流。

项目成果

Reiko Teshima: "Effect of an ectokinase inhibitorK252b on degranulatio and Ca^<2+> signals of RBL-2H3 cells and human basophils" J.Immunol.159. 964-969 (1997)

Reiko Teshima：“外激酶抑制剂K252b对RBL-2H3细胞和人嗜碱性粒细胞的脱颗粒和Ca 2+ 信号的影响”J.Immunol.159。

Reiko Teshima: "Effect of ectokinase inhibitor K252b on degranulatio and Ca^<2+> signals of RBL-2H3 cells and human basophils" J.Immunol.159. 964-969 (1997)

Reiko Teshima：“外切激酶抑制剂K252b对RBL-2H3细胞和人嗜碱性粒细胞的脱颗粒和Ca^2信号的影响”J.Immunol.159。

Mayumi Suzuki: "Soluble factors from Rat Basophilic Lenkemia (RBL-2H3) cells stimulated coopera tively the neurite out growth of PC12 cells" Biol.Pharm.Bull.21. 1267-1271 (1998)

Mayumi Suzuki：“来自大鼠嗜碱性粒细胞白血病 (RBL-2H3) 细胞的可溶性因子协同刺激 PC12 细胞的神经突生长”Biol.Pharm.Bull.21。

R.Teshima, Y.Saito, H.Ikebuchi, N.R.D.Silva, Y.Morita, M.Nakanishi, J.Sawada and S.Kitani: "Effect of an ecto-kinase inhibitor, K252b, on degranulation and Ca2+ signals of RBL-2H3 cells and human basophils" J.Immunology. 159. 964 (1997)

R.Teshima、Y.Saito、H.Ikebuchi、N.R.D.Silva、Y.Morita、M.Nakanishi、J.Sawada 和 S.Kitani：“外激酶抑制剂 K252b 对 RBL- 脱颗粒和 Ca2 信号的影响

Reiko Teshima: "Casein-Kinase-II-like ectokinase activity on RBL-2H3 cells" Immunol.Letter. (in press). (1999)

Reiko Teshima：“RBL-2H3 细胞上的酪蛋白激酶 II 样外切酶活性”Immunol.Letter。