Comparative analysis of involvement of UGT1 and UGT2 splice variants of UDP-galactose transporter in glycosylation of macromolecules in MDCK and CHO cell lines.
Comparative analysis of involvement of UGT1 and UGT2 splice variants of UDP-galactose transporter in glycosylation of macromolecules in MDCK and CHO cell lines.
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DOI：
10.1007/s10719-011-9348-z
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发表时间：
2011-10
影响因子：
3
通讯作者：
Olczak, Mariusz
中科院分区：
文献类型：
--
作者：
Maszczak-Seneczko, Dorota;Olczak, Teresa;Wunderlich, Livius;Olczak, Mariusz
核苷酸糖转运蛋白将核苷酸糖递送到高尔基体和内质网中。本研究旨在进一步表征MDCK和CHO细胞系中哺乳动物UDP-半乳糖转运蛋白（UGT）。MDCK-RCAr和CHO-Lec 8突变体细胞系在UGT转运蛋白中有缺陷，尽管它们表现出一定水平的半乳糖基化。以前，在两种细胞系中仅鉴定出单一形式的UGT，MDCK细胞中的UGT 1和CHO细胞中的UGT 2。我们已经在CHO（UGT 1）和MDCK（UGT 2）细胞中鉴定了第二UGT剪接变体。与UGT 1相比，UGT 2在几乎所有检查的哺乳动物组织和细胞系中更丰富，但MDCK细胞表现出两种剪接变体的不同相对分布。互补分析表明，这两个UGT剪接变异体是必要的N-和O-糖基化的蛋白质。与野生型细胞相比，两种突变细胞系产生的软骨素-4-硫酸酯水平仅略低。这种缺陷通过两种UGT剪接变体的过表达来纠正。MDCK-RCAr突变细胞不产生硫酸角质素，并且这种效应不被UGT剪接变体（单独或组合过表达）校正。在这里，我们证明了这两种UGT剪接变体对蛋白质的糖基化是重要的。与MDCK细胞相反，MDCK-RCAr突变细胞可能在硫酸角质素生物合成途径中具有额外的缺陷。本文的在线版本（doi：10.1007/s10719-011-9348-z）包含补充材料，可供授权用户使用。
Nucleotide sugar transporters deliver nucleotide sugars into the Golgi apparatus and endoplasmic reticulum. This study aimed to further characterize mammalian UDP-galactose transporter (UGT) in MDCK and CHO cell lines. MDCK-RCAr and CHO-Lec8 mutant cell lines are defective in UGT transporter, although they exhibit some level of galactosylation. Previously, only single forms of UGT were identified in both cell lines, UGT1 in MDCK cells and UGT2 in CHO cells. We have identified the second UGT splice variants in CHO (UGT1) and MDCK (UGT2) cells. Compared to UGT1, UGT2 is more abundant in nearly all examined mammalian tissues and cell lines, but MDCK cells exhibit different relative distribution of both splice variants. Complementation analysis demonstrated that both UGT splice variants are necessary for N- and O-glycosylation of proteins. Both mutant cell lines produce chondroitin-4-sulfate at only a slightly lower level compared to wild-type cells. This defect is corrected by overexpression of both UGT splice variants. MDCK-RCAr mutant cells do not produce keratan sulfate and this effect is not corrected by either UGT splice variant, overexpressed either singly or in combination. Here we demonstrate that both UGT splice variants are important for glycosylation of proteins. In contrast to MDCK cells, MDCK-RCAr mutant cells may possess an additional defect within the keratan sulfate biosynthesis pathway. The online version of this article (doi:10.1007/s10719-011-9348-z) contains supplementary material, which is available to authorized users.
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影响因子：
2.7
作者：
Ishida, N;Yoshioka, S;Kawakita, M
通讯作者：
Kawakita, M
DOI：
10.1073/pnas.0608159103
发表时间：
2006-10-31
影响因子：
11.1
作者：
Caffaro, Carolina E.;Hirschberg, Carlos B.;Berninsone, Patricia M.
通讯作者：
Berninsone, Patricia M.
影响因子：
4.8
作者：
Kawar, ZS;Haslam, SM;Cummings, RD
通讯作者：
Cummings, RD
影响因子：
4.8
作者：
Eckhardt, M;Gotza, B;Gerardy-Schahn, R
通讯作者：
Gerardy-Schahn, R
影响因子：
4.8
作者：
Oelmann, S;Stanley, P;Gerardy-Schahn, R
通讯作者：
Gerardy-Schahn, R