Targeted proteo-glycomics analysis of Sialyl Lewis X antigen expressing glycoproteins secreted by human hepatoma cell line
Targeted proteo-glycomics analysis of Sialyl Lewis X antigen expressing glycoproteins secreted by human hepatoma cell line
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DOI：
10.2116/analsci.19.85
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发表时间：
2003-01-01
影响因子：
1.6
通讯作者：
Matsumoto, K
中科院分区：
文献类型：
--
作者：
Koji, HT;Shibukawa, K;Matsumoto, K
唾液酸化路易斯X(sialyl Lewis X，SLEX)抗原Neu5Acalpha2-3Galbeta1-4(Fucalpha1-3)G1cNAc-R在细胞间相互作用中发挥重要作用：例如，E-和P-选择素介导表达SLEX的白细胞进入炎症区域。与HH-7细胞相比，人肝细胞癌细胞系HepG2在分泌糖蛋白和细胞表面高表达SLEX。采用双向聚丙烯酰胺凝胶电泳法和基质辅助激光解吸电离飞行时间质谱仪(MALDI-TOFMS)鉴定表达SLEX的糖蛋白，包括转铁蛋白、α、抗胰蛋白酶、α(2)-HS糖蛋白和β(2)-糖蛋白。我们用MALDI-TOFMS结合外切糖苷酶消化分析了这些糖蛋白的N-糖链；我们的结果表明多岩藻糖化和高支化的N-糖链增加。1，3-岩藻糖基转移酶在HepG2细胞中的高表达可能是糖蛋白中高α-1，3-岩藻糖基化的原因。
Sialyl Lewis X (SLEX) antigen, Neu5Acalpha2-3Galbeta1-4 (Fucalpha1-3) G1cNAc-R, plays important roles in cell-to-cell interaction: for example, the E- and P-selectin-mediated influx of SLEX expressing leukocytes into inflamed areas. A human hepatocellular carcinoma cell line, HepG2 cells, was highly expressed SLEX on secreted glycoproteins and cell surface, in contrast with HuH-7 cells. We identified SLEX expressing glycoproteins in HepG2 cultured medium by two-dimensional polyacrylamide gel electrophoresis, followed by in gel digestion and peptide mass fingerprint using matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOFMS), including transferrin, a,antitrypsin, alpha(2)-HS glycoprotein and beta(2)-glycoprotein. We analyzed N-glycans of these glycoproteins by MALDI-TOFMS in combination with exoglycosidase digestion; our results indicate increases in poly-fucosylated and high-branched N-glycans. High alpha1,3-fucosylation in glycoproteins would be caused by increased expression of alpha1,3-fucosyltransferase activities in HepG2 cells.