Structures and functions of the sugar chains of human leukocyte common antigen CD45

人白细胞共同抗原CD45糖链的结构和功能

• 批准号: 06680578
• 负责人: FURUKAWA Kiyoshi
• 金额: $ 1.41万
• 依托单位: Tokyo Metropolitan Institute of Gerontology
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for General Scientific Research (C)
• 财政年份: 1994
• 资助国家: 日本
• 起止时间: 1994 至 1995
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-06680578/
• 关键词: CD45; Asn-linked sugar chains; Sialylation; CD22; Mucin-type sugar chains; アスパラギン結合型糖鎖

CD45 is a leukocvte-specific cell surface glycoprotein with protein tyrosin phosphatase activity, and involved in the antigen-driven T cell growth.Recently B cell surface antigen CD22 has been shown to bind sialic acid on the sugar chains of T cell surface CD45.In order to eludate the function of the sugar chains of CD45, the carbohydrate structures of CD45 sugar chains were determined in the present study.(1) Structures of Asn-linked sugar chains : The sugar chains were released by hydrazinolysis from CD45 purified from human peripheral blood T cells, and reduced with NaB^3H_4.Carbohydrate structures of the radio-active sugar chains were determined by Bio-Gel P-4 column chromato-graphy in combination with exoglycosidase digestion, and by methylation analysis.The results showed that CD45 contains mainly tetra-and tri-antennary complex-type sugar chains with one to two sialic acid residues.(2) Structure of mucin-type sugar chains : The sugar chains were released by incubation of CD45 with alkaline-borohydride/borotritide solution.The redioactive sugar chains were analyzed as described above.The results showed that most sugar chains are neutral, and they are with core 1[Galbeta1-3GalNAc] and core 2 [Galbeta1-4GlcNAcbeta1-6(Galbeta1-3GalNAc)] structures.The sugar chain also contained i and l antigens, although their amounts are extremely small.(3)Binding assay of CD45 sugar chains to a CD22-immobilized column : The radioactive Asn-and mucin-type sugar chains thus prepared were applied to the column.The result showed that no sugar chains bind to the column, while heavily sialylated oligosaccharides prepared from fetuin and orosomucoid interact to.These results indicate that the sugar chains of CD45 from human peripheral blood T cells are not adequately sialylated for binding to CD22.

CD45是一种白细胞特异性细胞表面糖蛋白，具有蛋白酪氨酸磷酸酶活性，参与抗原驱动的T细胞生长。最近发现B细胞表面抗原CD22与T细胞表面糖链上的唾液酸结合。为了揭示CD45糖链的功能，本研究测定了CD45糖链的糖结构。(1)糖链结构：从人外周血T细胞提纯的CD45糖链通过肼分解释放，用Bio-Gel P-4柱层析结合外切糖苷酶消化和甲基化分析确定了放射性糖链的碳水化合物结构。结果表明，CD45主要含有四触角和三触角的复合型糖链，含有一到两个唾液酸残基。(2)粘蛋白糖链的结构：CD45与碱性硼氢化物/三硼三肽溶液孵育释放糖链。如上所述分析了折光糖链。结果表明，大多数糖链是中性的，糖链上还含有I和L抗原，但数量极少。(3)CD45糖链与CD22的结合分析：将制备的放射性天冬氨酸型和粘蛋白型糖链与CD22结合。结果表明，糖链上没有糖链与CD22结合，而由胎球蛋白和类粘液蛋白制备的重链寡糖相互作用。

项目成果

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