School of Pharmacy, Kinki University, Kowakae 3-4-1, Higashi-Osaka 577-8502, Japan.
J Pharm Biomed Anal. 2012 Nov;70:718-26. doi: 10.1016/j.jpba.2012.06.035. Epub 2012 Jul 4.
In the series of our previous reports, we showed that some cancer cell lines specifically express polylactosamine-type N-glycans and such glycans were often modified with fucose and sulfate residues. To confirm the proteins expressing these glycans, glycopeptide mixture obtained by digestion of whole protein fractions with trypsin was captured by a polylactosamine-specific lectin, Datura strasmonium agglutinin (DSA). And the peptides and glycans of the captured glycopeptides after digestion with N-glycoamidase F were extensively analyzed by HPLC and MS techniques. We found that some glycoproteins such as CD107a and CD107b commonly contained polylactosamine-type glycans in all the examined cancer cells. But integrin-α5 (CD49e) and carcinoembryonic antigen-related cell adhesion molecule 5 (CD66e) having these glycans were specifically found in U937 (human T-lymphoma) and MKN45 (human gastric cancer) cells, respectively. These data clearly indicate that specific glycans attached to specific proteins will be promising markers for specific tumors with high accuracy.
在我们之前的一系列报告中，我们表明一些癌细胞系特异性表达多糖型 N-糖基化，并且这些糖基通常被岩藻糖和硫酸基修饰。为了确认表达这些糖的蛋白质，用胰蛋白酶消化全蛋白级分获得的糖肽混合物被一种多乳糖胺特异性凝集素，颠茄凝集素（DSA）捕获。然后，用 N-糖苷酶 F 消化后捕获的糖肽中的肽和聚糖通过 HPLC 和 MS 技术进行广泛分析。我们发现，一些糖蛋白，如 CD107a 和 CD107b，在所有检查的癌细胞中通常都含有多糖型聚糖。但是，具有这些糖的整合素-α5（CD49e）和癌胚抗原相关细胞粘附分子 5（CD66e）分别特异性地存在于 U937（人 T 淋巴细胞瘤）和 MKN45（人胃癌）细胞中。这些数据清楚地表明，特异性地附着于特异性蛋白质的特异性聚糖将成为具有高精度的特异性肿瘤的有前途的标志物。
