Mucins and their O-Glycans from human bronchial epithelial cell cultures
Mucins and their O-Glycans from human bronchial epithelial cell cultures
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DOI：
10.1152/ajplung.00108.2004
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发表时间：
2004-10-01
影响因子：
4.9
通讯作者：
Davis, CW
中科院分区：
文献类型：
--
作者：
Holmén, JM;Karlsson, NG;Davis, CW
在阻塞性气道疾病中一个长期存在的问题是，所观察到的粘蛋白组成和/或翻译后糖基化的变化是由于遗传因素还是环境因素。我们测试了来自非囊性纤维化（CF）或CF患者的第二代原代人支气管上皮细胞培养物分泌的粘蛋白是否具有本质上不同的特异性粘蛋白组成，以及这些粘蛋白是否被不同地糖基化。CF和非CF培养物均主要产生MUC 5 B，如通过用粘蛋白特异性抗体的定量琼脂糖凝胶蛋白质印迹所判断的：MUC 5 B以比MUC 5AC高10倍的水平存在，这与我们先前的mRNA研究一致（Bernacki SH，纳尔逊AL，Abdullah L，Sheehan JK，Harris A，William DC，and Randell SH. Am J Respir Cell Mol Biol 20：595 - 604，1999）。从纯化的非CF和CF粘蛋白中释放的O-连接寡糖通过HPLC质谱法研究具有高度可变的聚糖结构，并且两组之间没有可观察到的差异。因此，在细胞培养的非感染/非炎症条件下，与CF表型相关的特异性粘蛋白或其O-聚糖没有差异。我们的结论是，直接从患者身上取样的粘蛋白中观察到的差异最有可能是由于与感染和/或炎症相关的环境因素。
A longstanding question in obstructive airway disease is whether observed changes in mucin composition and/or posttranslational glycosylation are due to genetic or to environmental factors. We tested whether the mucins secreted by second-passage primary human bronchial epithelial cell cultures derived from noncystic fibrosis (CF) or CF patients have intrinsically different specific mucin compositions, and whether these mucins are glycosylated differently. Both CF and non-CF cultures produced MUC5B, predominantly, as judged by quantitative agarose gel Western blots with mucin-specific antibodies: MUC5B was present at similar to10-fold higher levels than MUC5AC, consistent with our previous mRNA studies (Bernacki SH, Nelson AL, Abdullah L, Sheehan JK, Harris A, William DC, and Randell SH. Am J Respir Cell Mol Biol 20: 595 - 604, 1999). O-linked oligosaccharides released from purified non-CF and CF mucins and studied by HPLC mass spectrometry had highly variable glycan structures, and there were no observable differences between the two groups. Hence, there were no differences in either the specific mucins or their O-glycans that correlated with the CF phenotype under the noninfected/noninflammatory conditions of cell culture. We conclude that the differences observed in the mucins sampled directly from patients are most likely due to environmental factors relating to infection and/or inflammation.