Targeted glycoproteomic identification of cancer cell glycosylation.
Targeted glycoproteomic identification of cancer cell glycosylation.
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DOI：
10.1093/glycob/cwp065
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发表时间：
2009-08
期刊：
影响因子：
4.3
通讯作者：
Taylor ME
中科院分区：
文献类型：
--
作者：
Powlesland AS;Hitchen PG;Parry S;Graham SA;Barrio MM;Elola MT;Mordoh J;Dell A;Drickamer K;Taylor ME
GalMBP是一段血清甘露糖结合蛋白片段，已被修改为含半乳糖配体的探针。糖链阵列筛选显示，GalMBP的糖识别结构域选择性地结合常见的肿瘤相关糖链，包括Lewis结构和T抗原，这表明GalMBP等工程糖链结合蛋白是表征含有肿瘤相关糖链的糖蛋白的新工具。用放射性标记的GalMBP对MCF7乳腺癌细胞提取液和细胞膜的印迹结果表明，它与一组选定的高分子糖蛋白结合，这些高分子糖蛋白可以在用GalMBP构建的亲和柱上从MCF7细胞中纯化。这些糖蛋白的蛋白质组学和糖质谱分析表明，它们是CD98hc的形式，其糖链含有大量岩藻糖化的末端，包括leisX和lewy结构。该配体池被发现包括抗CD15抗体和内皮清道夫受体C型凝集素的靶配体，CD15抗体通常用于检测肿瘤上的Lewis X抗原，内皮清道夫受体C型凝集素可能通过与该抗原的相互作用参与肿瘤转移。对其他乳腺癌细胞株的调查显示，导致GalMBP结合的糖基化类型有很大的差异。更高水平的结合要么与各种不同细胞表面糖蛋白上携带的外臂岩藻糖化结构的存在有关，要么与携带T抗原的粘蛋白MUC1的高水平存在有关。
GalMBP is a fragment of serum mannose-binding protein that has been modified to create a probe for galactose-containing ligands. Glycan array screening demonstrated that the carbohydrate-recognition domain of GalMBP selectively binds common groups of tumor-associated glycans, including Lewis-type structures and T antigen, suggesting that engineered glycan-binding proteins such as GalMBP represent novel tools for the characterization of glycoproteins bearing tumor-associated glycans. Blotting of cell extracts and membranes from MCF7 breast cancer cells with radiolabeled GalMBP was used to demonstrate that it binds to a selected set of high molecular weight glycoproteins that could be purified from MCF7 cells on an affinity column constructed with GalMBP. Proteomic and glycomic analysis of these glycoproteins by mass spectrometry showed that they are forms of CD98hc that bear glycans displaying heavily fucosylated termini, including Lewisx and Lewisy structures. The pool of ligands was found to include the target ligands for anti-CD15 antibodies, which are commonly used to detect Lewisx antigen on tumors, and for the endothelial scavenger receptor C-type lectin, which may be involved in tumor metastasis through interactions with this antigen. A survey of additional breast cancer cell lines reveals that there is wide variation in the types of glycosylation that lead to binding of GalMBP. Higher levels of binding are associated either with the presence of outer-arm fucosylated structures carried on a variety of different cell surface glycoproteins or with the presence of high levels of the mucin MUC1 bearing T antigen.
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影响因子：
4.8
作者：
Kolatkar, AR;Weis, WI
通讯作者：
Weis, WI
影响因子：
4
作者：
Kobata, A;Amano, J
通讯作者：
Amano, J
DOI：
10.1073/pnas.96.2.371
发表时间：
1999-01-19
影响因子：
11.1
作者：
Ma, Y;Uemura, K;Kawasaki, T
通讯作者：
Kawasaki, T
影响因子：
4.8
作者：
Nakamura, E;Sato, M;Minato, N
通讯作者：
Minato, N
影响因子：
4.3
作者：
Coombs PJ;Taylor ME;Drickamer K
通讯作者：
Drickamer K