Synthesis and application of functional domain of macrophage scavenger receptor

巨噬细胞清道夫受体功能域的合成及应用

• 批准号: 10470494
• 负责人: DOI Takefumi
• 金额: $ 6.85万
• 依托单位: Osaka University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (B).
• 财政年份: 1998
• 资助国家: 日本
• 起止时间: 1998 至 2000
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-10470494/
• 关键词: macrophage; scavenger receptor; model molecule; aggregation; HSP70; HSP90; GAPDH; phosphorylation; 細胞質ドメイン; フコイダン; LPS; NO産生; アフィニティカラム; 機能ドメイン; ペプチド分子; 核酸分子

We have synthesized the ligand binding domain of macrophage scavenger receptor (MSR) and investigated the interaction of this molecule to polynucleotides and peptide fragments in apolipoprotein B-100. We found that a certain aggregate form of both polynucleotides and peptides modified by acetic acid or acrolein could compete the ligand binding of MSR.X-ray fiber diffraction analysis suggested that the competitive peptides modified by acrolein form a "cross β structure" as detected on amyloid β-peptide.We have also synthesized the cytoplasmic domain of MSR.Affinity column with this domain was used for isolating the molecules to associate with MSR.By this column, HSP70, HSP90, aminopeptidase, S-adenosylhomocysteinase, glyceraldehyde 3-phosphate dehydrogenase (GAPDH) and a unknown protein were isolated. We are now investigating the function of these proteins for signal transduction of MSR.We have analized the point mutants of which each serine or threonine residue was substituted by alanine in cytoplasmic domain of MSR.We found that the serine 16, 16th amino acid residue from N-terminus of MSR, was a residue to be phosphorylated by protein kinase and to play an important role for the signal transduction through MSR.

我们合成了巨噬细胞清道夫受体（MSR）的配体结合域，并研究了该分子与载脂蛋白B-100中多核苷酸和肽片段的相互作用。我们发现，由乙酸或丙烯醛修饰的多核苷酸和多肽的某种聚集形式可以竞争MSR的配体结合。x射线纤维衍射分析表明，经丙烯醛修饰的竞争肽在淀粉样β-肽上呈“交叉β结构”。我们还合成了MSR的细胞质结构域。利用具有该结构域的亲和柱分离与MSR相关的分子。通过该柱分离到HSP70、HSP90、氨基肽酶、s -腺苷高半胱氨酸酶、甘油醛3-磷酸脱氢酶（GAPDH）和未知蛋白。我们现在正在研究这些蛋白在MSR信号转导中的功能。我们分析了MSR细胞质区域中每个丝氨酸或苏氨酸残基被丙氨酸取代的点突变体。我们发现丝氨酸16 （MSR n端第16个氨基酸残基）是一个被蛋白激酶磷酸化的残基，在MSR的信号转导中起重要作用。

项目成果

H.Shirai: "Structure and function of type I and II macrophage scavenger receptors."Mech.Ageing Dev.. 111. 107-121 (1999)

H.Shirai：“I 型和 II 型巨噬细胞清道夫受体的结构和功能。”Mech.Ageing Dev.. 111. 107-121 (1999)

Y.Yamada, T.Doi, T.Hamakubo, & T.Kodama: "Scavenger receptor family proteins : roles for atherosclerosis, host defense and disorders of the central nervous system."Cell.Mol.Life Sci.. 54. 628-640 (1998)

Y.山田、T.土井、T.滨久保、

Y.Yamada: "Scavenger receptor family proteins : roles for atherosclerosis ; host defence and disorders of the central nervous system."Cell.Mol.Life Sci.. 54. 628-640 (1998)

Y.Yamada：“清道夫受体家族蛋白：动脉粥样硬化的作用；宿主防御和中枢神经系统疾病。”Cell.Mol.Life Sci.. 54. 628-640 (1998)

Y.Takakura: "Characterization of plasmid DNA binding and uptake by peritoneal macrophages from class A scavenger receptor knockout mice."Pharmaceutical Research. 16. 503-508 (1999)

Y.Takakura：“A 类清道夫受体敲除小鼠腹腔巨噬细胞的质粒 DNA 结合和摄取的表征。”药物研究。

W.Yu, A.Shimoyama, T.Uneda, S.Obika, K.Miyashita, T.Doi, & T.Imanishi: "Gene transfer mediated by YKS-220 cationic particles : convenient and efficient gene delivery reagent."J.Biochem., (Tokyo). 125. 1034-1038 (1999)

W.Yu、A.Shimoyama、T.Uneda、S.Obika、K.Miyashita、T.Doi、