AN IDENTIFICATION AND CHARACTERIZATION OF MENBRANE PROTEIN ASSOCIATED WITH CELL ACTIVATIONS LY BACTERIAL LIPOPOLYSACCHARIDES

与细胞活化相关的细菌脂多糖膜蛋白的鉴定和表征

• 批准号: 11670270
• 负责人: KIRIKAE Teruo
• 金额: $ 2.24万
• 依托单位: INTERNATIONAL MEDICAL CENTER OF JAPAN
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 1999
• 资助国家: 日本
• 起止时间: 1999 至 2000
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-11670270/
• 关键词: LPS; LIPOPOLYSACCHARIDE; ENDOTOXIN; LIPIDA; MACROPHAGE; PE-406; PE-506; STOROMA-DERIVED CELL; Lipoplysaccharide; LPS結合タンパク質; タキソール

A bacterial endotoxin lipopolysaccharide (LPS) is known to induce the expression of various cytokines via activation of host cells, including macrophages. To identify and characterize the novel molecules which play integral roles in the LPS-induced cell activation, we utilized antibodies against membrane proteins and isotopically labeled lipid A, and detected some candidates. We have previously shown that murine stroma-derived ST2 cells do not express CD14, and have approximately the same LPS responsiveness as macrophages. We here raised polyclonal antibodies against purified cell membrane from ST2 cells, as well as LPS binding protein partially purified from the ST2 cell membrane fraction by HPLC.Expression of particular proteins in the macrophage membrane was observed by concentration, separation and visualization of macrophage membrane proteins by immunoprecipitation and subsequent Western blot with antibodies raised against the membrane fraction. We also detected 57 kDa and 53kDa proteins as lipid A binding proteins by ligand blot, respectively with [^3H]-lipid A precursor (PE-406) and [^3H]-lipid A (PE-506).

已知细菌内毒素脂多糖（LPS）通过激活宿主细胞（包括巨噬细胞）诱导各种细胞因子的表达。为了识别和表征在LPS诱导的细胞激活中起着不可或缺的作用的新型分子，我们利用针对膜蛋白和同位素标记的脂质A的抗体，并检测到一些候选物。我们先前已经表明，鼠基质衍生的ST2细胞不表达CD14，并且与巨噬细胞的LPS响应性大致相同。我们在这里提出了来自ST2细胞纯化的细胞膜的多克隆抗体，以及通过HPLC通过HPLC表达巨噬细胞膜中特定蛋白质在ST2细胞膜分数中部分纯化的LPS结合蛋白，通过浓度与巨噬细胞蛋白质的浓度和可视化的巨型膜分离和可视化观察到巨噬细胞膜中的特定蛋白质，并通过免疫膜蛋白的分离和可视化观察到。 分数。我们还与[^3H] -lipid A前体（PE-406）和[^3H] -lipid A（PE-506）一起，分别检测到57 kDa和53kDa蛋白作为脂质A的结合蛋白。

项目成果

Swierzko, A.S., Kirikae, T., kirikae, F., Hirata, M., et al.: "Biological activities of Proteus lipopolysaccharides and their interaction with polymyxin B and a 18-kilodalton cationic antimicrobial protein (CAP18) -derived peptide."J.Med. Microbiol.. 49.

Swierzko, A.S.、Kirikae, T.、kirikae, F.、Hirata, M. 等人：“变形杆菌脂多糖的生物活性及其与多粘菌素 B 和 18 千道尔顿阳离子抗菌蛋白 (CAP18) 衍生肽的相互作用。

Hashimoto,M.,Kirikae,F.Toyooka,K.,Kaneko,A., et al.: "Protective Effect of OK-432 on Mice against Endotoxemia and Infection with Pseudomonas aeruginosa and Salmonella enteritidis."Microbiol.Immunol.. (in press).

Hashimoto,M.、Kirikae,F.Toyooka,K.、Kaneko,A. 等人：“OK-432 对小鼠内毒素血症和铜绿假单胞菌和肠炎沙门氏菌感染的保护作用。”Microbiol.Immunol..（

Kawano, C., Muroi, K., Yoshizaka, M., Kirikae, T., Ozawa, K., et al.: "Filamented Pseudomonas aeruginosa and a markedly high level of endotoxin in cerebrospinal fluid"J.Infect.. 41. 114-115 (2000)

Kawano, C.、Muroi, K.、Yoshizaka, M.、Kirikae, T.、Ozawa, K.等人：“丝状铜绿假单胞菌和脑脊液中内毒素水平显着升高”J.Infect.. 41

"Important role of membrane-associated CD14 in the induction of IFN-g and subsequent nitric oxide production by murine macrophages in response to bacterial lipopolysaccharide."Eur. J.Biochem.. 267. 37-45 (2000)

“膜相关 CD14 在诱导 IFN-g 以及随后小鼠巨噬细胞响应细菌脂多糖产生一氧化氮中的重要作用。”Eur。

Swierzko,,A.S.,Kirikae,T.,Kirikae,F.,Hirata,M., et al.: "Biological activities of Proteus lipopolysaccharides and their interaction with polymyxin B and a 18-kilodalton cationic antimicrobial protein (CAP18)-derived peptide."J.Med.Microbiol.. 49. 127-138

Swierzko,,A.S.、Kirikae,T.、Kirikae,F.、Hirata,M. 等人：“变形杆菌脂多糖的生物活性及其与多粘菌素 B 和 18 千道尔顿阳离子抗菌蛋白 (CAP18) 衍生肽的相互作用