Alteration of site-directed sugar chains of α_1-acid glycoprotein in serum of patients and its clinical significance

患者血清中α_1-酸性糖蛋白定点糖链的改变及其临床意义

• 批准号: 13672433
• 负责人: MATSUMOTO Kojiro
• 金额: $ 1.47万
• 依托单位: Toho University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 2001
• 资助国家: 日本
• 起止时间: 2001 至 2003
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-13672433/
• 关键词: α_1-acid glycoprotein; N-glycans; α1-3 fucosylation; sialyl Lewis X antigen; MALDI-TOFMS; inflammation; α_1-酸性糖蛋白; 糖鎖; サイトカイン; α1-酸性糖蛋白; 糖鎖結合部位特異的糖鎖構造; 糖尿病

N-Glycans of α_1-acid glycoprotein (AGP) in sera of healthy individuals and patients with inflammation were comparatively studied (Clin Chim Acta, 2003). N-Glycans released from purified AGP with N-glycanase were treated with sialidase and subjected to MALDI-TOFMS analysis. N-Glycans of both groups were composed with bi-, tri-and tetra-antennary complex-types and the contents of α1-3 fucosylated N-glycans were bi-□tri-<tetra-antennary. In inflammation patients, increases in bi-antennary, decreases in tri-and tetra-antennary and increases in α1-3 fucosylation were significant.N-Glycans of 5 glycosylation sites in AGP were separately compared (in preparation). Purified AGP was treated with endopeptidase Glu-C and N-glycans of each glycopeptides isolated by a reversed phase HPLC were determined by MALDI-TOFMS. Each glycosylation site was composed with different and characteristic antennary glycans, bi-and tri-in sites 1 and 2 and tri-and tetra-antennary in sites 3 to 5. In inflammatory, i … More ncreases in bi-antennary, decreases in tri-and tetra-antennary and increases in α_1-3 fucosylation were commonly observed in all glycosylation sites, in larger degrees in acute than chronic inflammation.To determine cancerous alteration of N-glycans, N-glycans of glycoproteins secreted from human hepatoma cell lines HuH7 and HepG2 were studied (Anal Sci, 2003). Increases in tri-and tetra-antennary and α1-3 fucosylation were prominently detected in several glycoproteins secreted by HepG2 cells. Moreover, α1-3 fucosylation activities toward NeuAcα2-3Galβ1-4GlcNAc-R were detected 20-fold highly in HepG2, comparing to HuH7.To clarify the biological roles of high-fucosylated N-glycans, NK cells were stimulated on plates coated with high-fucosylated glycoproteins purified from HepG2 culture, medium (unpublished result). High-fucosylated glycoproteins will modulate the NK cell functions, since protein with 17kDa was stimulated to phosphorylate on its tyrosine residues. Further study is in progress. Less

比较研究了健康人与炎症患者血清中α_1-酸性糖蛋白（AGP）的n -聚糖（clinchinacta, 2003）。用唾液酸酶处理纯化AGP后释放的n -聚糖，并进行MALDI-TOFMS分析。两组的n -聚糖均由双、三、四天线络合物组成，α - 1-3浓缩的n -聚糖含量为双、三、四天线络合物。在炎症患者中，双触角蛋白增加，三、四触角蛋白减少，α - 1-3聚焦化显著增加。分别比较AGP中5个糖基化位点的n -聚糖（制备中）。用内肽酶Glu-C对纯化的AGP进行处理，反相高效液相色谱分离得到的每个糖肽的n -聚糖用MALDI-TOFMS测定。每个糖基化位点由不同的和特征的天线聚糖组成，二、三链位点1和2，三、四链位点3至5。在所有糖基化位点均可见到更多的双触角蛋白增加，三、四触角蛋白减少，α_1-3聚焦化增加，急性炎症的程度大于慢性炎症。为了确定n -聚糖的癌变，研究了人肝癌细胞系HuH7和HepG2分泌的糖蛋白的n -聚糖（肛门科学，2003）。HepG2细胞分泌的几种糖蛋白中，三、四天线和α - 1-3聚焦化显著增加。此外，HepG2对NeuAcα2-3Galβ1-4GlcNAc-R的α1-3聚焦活性比HuH7高20倍。为了阐明高度聚焦的n -聚糖的生物学作用，我们将NK细胞刺激在从HepG2培养基中纯化的高度聚焦的糖蛋白包被的板上（未发表的结果）。高度集中的糖蛋白会调节NK细胞的功能，因为含有17kDa的蛋白被刺激磷酸化其酪氨酸残基。进一步的研究正在进行中。少

项目成果

S Muto, T Takada, K Matsumoto: "Biological activities of human mannose-binding lecthin bound to two different ligand sugar structures, Lewis A and Lewis B antigens and high-mannose oligosaccharides."Biochim Biophys Acta. 1527(1-2). 39-46 (2001)

S Muto、T Takada、K Matsumoto：“人甘露糖结合凝集素与两种不同的配体糖结构（Lewis A 和 Lewis B 抗原以及高甘露糖寡糖）结合的生物活性。”Biochim Biophys Acta。

A Taniguchi, T Morishuina, Y Tsujita, Y Matsumoto, K Matsumoto: "Genomic structure, expression and transcriptional regulation of human Galβ1,3GalNAc α2,3-sialyltransferase gene."Biochem Biophys Res Commun. 300(2). 570-576 (2003)

A Taniguchi、T Morishuina、Y Tsujita、Y Matsumoto、K Matsumoto：“人类 Galβ1,3GalNAc α2,3-唾液酸转移酶基因的基因组结构、表达和转录调控。”Biochem Biophys Res Commun. 570(2)。 2003）

L Xu, Y Kurusu, K Takizawa, J Tanaka, K Matsumoto, A Taniguchi: "Transcriptional regulation of human β-galactoside α2,6-sialyltrans-ferase (hST6Gal I) gene in colon adenocarcinoma cell line."Biochem Biophys Res Commun. 307(4). 1070-1074 (2003)

L Xu、Y Kurusu、K Takizawa、J Tanaka、K Matsumoto、A Taniguchi：“结肠腺癌细胞系中人 β-半乳糖苷 α2,6-唾液酸转移酶 (hST6Gal I) 基因的转录调节。”Biochem Biophys Res Commun。 307（4）。1070-1074（2003）。

K Higai, K Shibukawa, S Muto, K Matsumoto: "Targeted proteo-glycomics analysis of sialyl Lewis X antigen expressing glycoproteins secreted by human hepatoma cell line"Anal Sci. 19(1). 85-92 (2003)

K Higai、K Shibukawa、S Muto、K Matsumoto：“对表达人肝癌细胞系分泌的唾液酸路易斯 X 抗原的糖蛋白进行靶向蛋白糖组学分析”Anal Sci。

Y Azuma, M Sakanashi, K Matsumoto: "The effect of α2,6-linked sialic acid on anti-IgM antibody-induced apoptosis in Karaos cells"Glycoconj J. 18(5). 419-424 (2001)

Y Azuma、M Sakanashi、K Matsumoto：“α2,6-连接唾液酸对抗 IgM 抗体诱导的 Karaos 细胞凋亡的影响”Glycoconj J. 18(5) (2001)。