Analysis of specific protein phosphorylation after infection with bacteria

细菌感染后特异性蛋白质磷酸化分析

• 批准号: 05454194
• 负责人: NAKANO Masayasu
• 金额: $ 5.12万
• 依托单位: Jichi Medical School
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for General Scientific Research (B)
• 财政年份: 1993
• 资助国家: 日本
• 起止时间: 1993 至 1994
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-05454194/
• 关键词: BACTERIAL INFECTION; PROTEIN PHOSPHORYLATION; SALMONELLA; ENDOTOXIN; LIPOPOLYSACCHARIDE; MACROPHAGE; リポ多糖

When murine peritoneal macrophages were cultured in vitro and infected with bacteria such as Salmonella typhimurium, S.enteritidis and others, a set of phosphorylated proteins appeared inthe cells. Some of the proteins, such as 65 kDa(pp65), are specific to bacterial lipipolysaccharide (LPS), while several others, such as 85 kDa (pp85) and 72 kDa (pp72) are specific to the live Salmonella infection. Cytochalacin D-treatment inhibited partially the phagocytosis oforganisms, but it did not inhibit the phosporylations of pp85 and pp72. When intracellular growth of infected Salmonella was inhibited by chloram-phenicol or nalidixic acid, the phosphorylation did not appear. These findings suggest that the phosphorylation is due to the intracellular growth of live organisms rather than phagocytic process. The phosphorylation of pp85 and pp72 was not observed by the infections with Mycobacterium bovis BCG,Legionella pneumophila, Mycobacterium intracellulare, Stapylococcus aureus, Listeria monocytogenes, Pseudomonas aeruginosa.Many phosphorylated proteins were observed by stimulation with LPS or heat-killed gram negative organisms. pp65 was the most dominant one in cytoplasm of macrophages. Amino acid sequences of pp65 was determined. It was a substrate of serine-kinase and a member of plastin family.

小鼠腹膜巨噬细胞在体外培养并感染鼠伤寒沙门氏菌、肠炎沙门氏菌等细菌时，细胞内出现一组磷酸化蛋白。其中一些蛋白质，如65 kDa(Pp65)是细菌脂多糖(LPS)所特有的，而另一些蛋白质，如85 kDa(Pp85)和72 kDa(Pp72)是活体沙门氏菌感染所特有的。胞查拉星D处理部分抑制了组织的吞噬作用，但不抑制pp85和pp72的磷酸化。当感染沙门氏菌的细胞内生长被氯霉素或奈利地酸抑制时，没有出现磷酸化。这些发现表明，磷酸化是由于活生物的细胞内生长，而不是吞噬过程。卡介苗、嗜肺军团菌、胞内分枝杆菌、金黄色葡萄球菌、单核细胞增生性李斯特菌、铜绿假单胞菌等感染均未观察到pp85和pp72的磷酸化。巨噬细胞胞浆中以pp65为主。测定了pp65的氨基酸序列。它是丝氨酸激酶的底物，是纤溶酶家族的成员。

项目成果

Shinomiya, H., Hirata, H., Saito, S., Yagisawa, H., and Nakano, M.: "Identification of the 65-kDa phosphoprotein in murine macrophages as a novel protein." Biochemical and Biophysical Research Communications. 202. 1631-1638 (1994)

Shinomiya, H.、Hirata, H.、Saito, S.、Yagisawa, H. 和 Nakano, M.：“鉴定小鼠巨噬细胞中的 65-kDa 磷蛋白是一种新型蛋白质。”

Saito,S.: "Protein phosphorylation in murine peritoneal macrophagec infected by infection with Salmonella species." Infection and Immunity. 62. 1551-1556 (1994)

Saito,S.：“受沙门氏菌感染的小鼠腹膜巨噬细胞中的蛋白质磷酸化。”

Nakano, M., Yamasu, H., Terada.Y., Shinomiya, H., and Saito, S.: "LPS-induced protein phosphorylation and monokine production in calcium ionophore-stimulated or BCG-infected C3H/HeJ macrophages." Bacterial Endotoxin : Recognition and Effetor Mechanisms (E

Nakano, M.、Yamasu, H.、Terada.Y.、Shinomiya, H. 和 Saito, S.：“钙离子载体刺激或 BCG 感染的 C3H/HeJ 巨噬细胞中 LPS 诱导的蛋白质磷酸化和单核因子产生。”

斉藤 慎二: "LPS刺激マクロファージのサイトカイン産生系へのプロテインカイネースの関与" 第41回毒素シンポジウム予稿集. 41. 57-59 (1994)

Shinji Saito：“LPS 刺激的巨噬细胞的细胞因子产生系统中蛋白激酶的参与”第 41 届毒素研讨会论文集 41. 57-59 (1994)。

中野昌康 他: "細菌内毒素によるマクロファージ内シグナル伝達とその制御" 医学微生物学の新しい展開1993. 198-206 (1993)

Masayasu Nakano等：“巨噬细胞内信号转导及其细菌内毒素的调节”医学微生物学新进展1993。198-206（1993）