Functional sialylated O-glycan to platelet aggregation on Aggrus (T1α/podoplanin) molecules expressed in Chinese hamster ovary cells
Functional sialylated O-glycan to platelet aggregation on Aggrus (T1α/podoplanin) molecules expressed in Chinese hamster ovary cells
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DOI：
10.1074/jbc.m407210200
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发表时间：
2004-09-10
影响因子：
4.8
通讯作者：
Osawa, M
中科院分区：
文献类型：
--
作者：
Kaneko, M;Kato, Y;Osawa, M
聚集蛋白，也称为T1 α和podoplanin，是一种新的血小板聚集诱导因子，在各种癌细胞中表达。已知聚集蛋白/T1 α/podoplanin在肺I型肺泡细胞或淋巴管内皮细胞中表达。然而，其生理作用尚未阐明。为了评估糖基化对聚集血小板聚集活性的归因，在一系列中国仓鼠卵巢（CHO）细胞突变体中稳定表达重组分子，这些突变体包括N-聚糖缺陷型Lec 1、CMP-唾液酸转运蛋白缺陷型Lec 2和UDP-半乳糖转运蛋白缺陷型Lec 8。建立了一种新的抗人Aggrus单克隆抗体YM-1，用于检测人Aggrus在CHO细胞突变体上的表达。凝集素对Lec 1细胞有诱导血小板聚集作用，而对Lec 2和Lec 8细胞无诱导作用。此外，通过凝集素印迹分析聚集体上的聚糖。在CHO和Lec 1细胞中表达的Aggrus显示小麦胚芽凝集素、Jacalin和野豌豆凝集素结合。凝集素印迹结果表明，唾液酸化的核心1结构，唾液酸加上Galbeta 1，3GalNAc-Ser/Thr，是血小板聚集活性的关键。这种寡糖结构被称为肿瘤相关抗原，其与癌细胞的转移过程潜在相关。
Aggrus, also called T1alpha and podoplanin, is a novel platelet aggregation-inducing factor that is expressed in various carcinoma cells. Aggrus/T1alpha/podoplanin is known to be expressed in lung type I alveolar cells or lymphatic endothelial cells. However, its physiological role has not been clarified. To assess the attribution of glycosylation to Aggrus platelet aggregation activity, recombinant molecules were stably expressed in a series of Chinese hamster ovary (CHO) cell mutants, N-glycan-deficient Lec1, CMP-sialic acid transporter-deficient Lec2, and UDP-galactose transporter-deficient Lec8. A new anti-human Aggrus monoclonal antibody, YM-1, was established to detect the expression of human Aggrus on these CHO cell mutants. Aggrus on Lec1 cells induced platelet aggregation, but those on Lec2 and Lec8 cells did not. Further, the glycans on Aggrus were analyzed by lectin blotting. Aggrus expressed in CHO and Lec1 cells showed Wheat-germ agglutinin, Jacalin, and Vicia villosa lectin bindings. Lectin blotting results indicated that sialylated core 1 structures, sialic acid plus Galbeta1,3GalNAc-Ser/Thr, were critical for the platelet aggregation activity. This oligosaccharide structure is known as tumor-associated antigen, which is potentially related to the metastasis process of cancer cells.