Surface glycoproteomic analysis of hepatocellular carcinoma cells by affinity enrichment and mass spectrometric identification
Surface glycoproteomic analysis of hepatocellular carcinoma cells by affinity enrichment and mass spectrometric identification
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通过亲和富集和质谱鉴定对肝细胞癌细胞进行表面糖蛋白组学分析
DOI：
10.1007/s10719-012-9420-3
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发表时间：
2012-08-01
影响因子：
3
通讯作者：
Qian, Xiaohong
中科院分区：
文献类型：
--
作者：
Mi, Wei;Jia, Wei;Qian, Xiaohong
细胞表面糖蛋白是与恶性生长相关的最常见的现象之一。肝细胞癌是世界上最严重的恶性肿瘤之一。大多数肝细胞癌细胞表面蛋白在肿瘤侵袭转移过程中通过糖基化修饰。因此，细胞表面糖蛋白的鉴定可以为肝癌的诊断和治疗提供重要的信息，也是潜在的肝细胞癌诊断生物标志物和治疗靶点的潜在来源。然而，由于肝细胞癌细胞表面糖蛋白的疏水性、低丰度和低溶解性，蛋白质组学方法很少对其进行鉴定。近年来发展起来的细胞表面捕获技术(CSC)是一种针对膜糖蛋白的方法，它通过在完整的细胞表面进行糖链生物素化标记来亲和捕获膜糖蛋白。为了研究肝癌细胞表面糖蛋白质组的特征，探讨肝癌细胞侵袭转移的机制，我们对细胞表面捕获策略进行了改进和评价，并将其应用于肝癌细胞的表面糖蛋白质组分析。总共鉴定了116个独特的糖肽上的119个糖基化位点，对应于79种不同的蛋白质。其中，65个(54.6%)新的预测糖基化位点被确定为以前没有通过实验确定的。在鉴定的糖蛋白中，通过数据库搜索，82%被归类为膜蛋白，68%具有跨膜结构域(TMD)，24%被预测含有2-13个TMD。此外，在肝细胞癌细胞膜糖蛋白中检测到26个CD抗原和50个糖肽，占已鉴定糖肽总数的43%。其中许多已确定的糖蛋白与癌症有关，如CD44、CD147和EGFR。这是对肝细胞癌细胞表面糖蛋白的系统表征。本研究中鉴定的膜糖蛋白为探讨肝癌侵袭转移机制提供了非常有用的信息。
Cell surface glycoproteins are one of the most frequently observed phenomena correlated with malignant growth. Hepatocellular carcinoma (HCC) is one of the most malignant tumors in the world. The majority of hepatocellular carcinoma cell surface proteins are modified by glycosylation in the process of tumor invasion and metastasis. Therefore, characterization of cell surface glycoproteins can provide important information for diagnosis and treatment of liver cancer, and also represent a promising source of potential diagnostic biomarkers and therapeutic targets for hepatocellular carcinoma. However, cell surface glycoproteins of HCC have been seldom identified by proteomics approaches because of their hydrophobic nature, poor solubility, and low abundance. The recently developed cell surface-capturing (CSC) technique was an approach specifically targeted at membrane glycoproteins involving the affinity capture of membrane glycoproteins using glycan biotinylation labeling on intact cell surfaces. To characterize the cell surface glycoproteome and probe the mechanism of tumor invasion and metastasis of HCC, we have modified and evaluated the cell surface-capturing strategy, and applied it for surface glycoproteomic analysis of hepatocellular carcinoma cells. In total, 119 glycosylation sites on 116 unique glycopeptides were identified, corresponding to 79 different protein species. Of these, 65 (54.6 %) new predicted glycosylation sites were identified that had not previously been determined experimentally. Among the identified glycoproteins, 82 % were classified as membrane proteins by a database search, 68 % had transmembrane domains (TMDs), and 24 % were predicted to contain 2-13 TMDs. Moreover, a total of 26 CD antigens with 50 glycopeptides were detected in the membrane glycoproteins of hepatocellular carcinoma cells, comprising 43 % of the total glycopeptides identified. Many of these identified glycoproteins are associated with cancer such as CD44, CD147 and EGFR. This is a systematic characterization of cell surface glycoproteins of HCC. The membrane glycoproteins identified in this study provide very useful information for probing the mechanism of liver cancer invasion and metastasis.