A search with Pertussis Toxin for Cellular Factors Affecting the Properties of GTP-Binding Protein

用百日咳毒素寻找影响 GTP 结合蛋白特性的细胞因素

• 批准号: 04680184
• 负责人: HAZEKI Osamu
• 金额: $ 1.28万
• 依托单位: University of Tokyo
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for General Scientific Research (C)
• 财政年份: 1992
• 资助国家: 日本
• 起止时间: 1992 至 1993
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-04680184/
• 关键词: PERTUSSIS TOXIN; ADP-RIBOSYLATION; GTP-BINDING PROTEIN; LIPOPOLYSACCHARIDE; NEUTROPHILS; モノADPリボシル化; モノADアリボシル化; ザイモザン

Pertussis toxin-catalyzed ADP-ribosylation of membrane Gi (inhibitory guanine nucleotide-binding protein) was examined on the hypothesis that the rate of modification may reflect the membrane environment of Gi. Incubation of saponin-permeabilized neutrophils with activated pertussis toxin and NAD led to rapid ADP-ribosylation of cellular Gi. The rate of ADP-ribosylation was found to be decreased by prior treatment of the cells with lipopolysaccharide (LPS), in spite that the cellular content of Gi was unaffected by the treatment. Receptor-mediated activation of Gi by formyl-peptide caused no change in the rate of the modification. LPS potentiated the formyl-peptide-induced activation of phospholipase C and increase in intracellular Ca^<++> in intact neutrophils. The changes were accompanied by the increased GDP-GTP exchange and GTP-hydrolyzing activities of Gi in membrane preparations. These results suggest that LPS-treatment of neutrophils causes Gi quantitative change or altered interaction with other cellular component, which affects the rate of pertussis toxin-catalyzed ADP-ribosylation.

研究了百日咳毒素催化的膜抑制鸟嘌呤核苷酸结合蛋白(GI)的ADP-核糖化，其修饰速率可能反映了GI的膜环境。用激活的百日咳毒素和NAD孵育皂素渗透性中性粒细胞，导致细胞GI的快速ADP-核糖化。用脂多糖(LPS)预先处理细胞后，ADP-核糖化速率降低，但细胞内Gi的含量不受影响。受体介导的胃肠道甲酰基肽激活不改变修饰的速率。脂多糖可增强甲酰肽诱导的磷脂酶C的激活，并使完整中性粒细胞内钙离子水平升高。这些变化伴随着膜制剂中GI的GDP-GTP交换和GTP水解酶活性的增加。这些结果表明，内毒素处理中性粒细胞引起胃肠道数量的改变或改变与其他细胞成分的相互作用，从而影响百日咳毒素催化的ADP-核糖化的速率。

项目成果

Okada, T., Kawano, Y., Sakakibara, T., Hazeki, O.and Ui, M.: "Essential role of phosphatidylinositol 3-kinase in insulin-induced glucose transport and antilipolysis in rat adpiocytes. Studies with a selective inhibitor wortmannin." J.Biol.Chem.269. 3568-3

Okada, T.、Kawano, Y.、Sakakibara, T.、Hazeki, O. 和 Ui, M.：“磷脂酰肌醇 3-激酶在大鼠脂肪细胞中胰岛素诱导的葡萄糖转运和抗脂解作用中的重要作用。选择性抑制剂的研究

YATOMI,Y.: "Suppression by wortmannin of plalelet responses to stimuli due to inhibition of pleckstrin phosphorylation" Biochem.J.285. 745-751 (1992)

YATOMI，Y.：“由于抑制普莱克斯特林磷酸化，渥曼青霉素抑制血小板对刺激的反应”Biochem.J.285。

Okada,T.et al.: "Essential role of phosphatidylinositol 3-kinase in insulin-induced glucose transport and anti-lipolysis in rat adipocytes.Studies with a selective inhibitor wortmannin." J.Biol.Chem.269. 3568-3573 (1994)

Okada,T.等人：“磷脂酰肌醇 3-激酶在大鼠脂肪细胞中胰岛素诱导的葡萄糖转运和抗脂肪分解中的重要作用。选择性抑制剂渥曼青霉素的研究。”

Okada,T.: "Blockage of Chemotactic Peptide-induced Stimulation of Neutrophils by Wortmannin as a Result of Selectiue Inhibition of Phosphatidylinositol 3-Kinase" J.Biol.Chem.269. 3563-3567 (1994)

Okada,T.：“选择性抑制磷脂酰肌醇 3-激酶，阻断渥曼青霉素诱导的趋化肽诱导的中性粒细胞刺激”J.Biol.Chem.269。

櫨木修,宇井理生: "細菌毒素とADPリボシル化" 実験医学. 11. 245-249 (1994)

Osamu Kashigi，Rio Ui：“细菌毒素和 ADP 核糖基化”实验医学。 11. 245-249 (1994)