Structural variations in the glycosaminoglycan-protein linkage region of recombinant decorin expressed in Chinese hamster ovary cells
Structural variations in the glycosaminoglycan-protein linkage region of recombinant decorin expressed in Chinese hamster ovary cells
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DOI：
10.1093/glycob/7.8.1175
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发表时间：
1997-12-01
期刊：
影响因子：
4.3
通讯作者：
Sugahara, K
中科院分区：
文献类型：
--
作者：
Kitagawa, H;Oyama, M;Sugahara, K
Decorin是一种小分子成纤维细胞蛋白多糖，由一个核心蛋白和一个单一的软骨素/皮肤素硫酸盐链组成，研究了在中国仓鼠卵巢细胞中表达的重组Decorin的糖-蛋白连接区结构，并在培养上清液中分泌，经阴离子交换层析分离。用碱性NaBH4消除粘胶蛋白中的糖胺多糖链，然后用软骨素酶ABC消化。这些处理得到了来自糖-蛋白质连接区的主要和少量的六糖糖醛二醇，用酶切结合高效液相色谱分析了它们的结构，其中两个化合物具有传统的六糖核心，Delta六糖α1-3GalNAcβ1-4GlcAβ1-3GalNAcβ1-3GalNAcβ1-4Xyl-ol。一种是非硫化的，另一种是在GalNAc残基的C4上单硫化的，它们分别占总连接区的12%和60%，另一种化合物具有六糖糖醛二醇，其内部含有艾杜糖醛酸残基Delta HEXAα1->；3GalNAc(4-硫酸盐)β1-4IdoAα1-3Galβ1-3Galβ1-3Galβ1-4Xyl-ol，这是先前从牛主动脉的皮肤素硫酸盐蛋白多糖(Sugahara et al.，J.Biol)中分离的五个连接的六糖糖醇中的一个。化学270,7204-7212,1995)，该化合物占总连接区的11%，核心蛋白的连接六糖区域的这些结构变化与人α-胰蛋白酶抑制物(Yamada ef al.，GlycoBiology，5,335-341,1995)和尿胰蛋白酶抑制物(Yamada ef al.，EUR，J.Biochem，233,687-693,1995)的连接六糖结构的一致性形成鲜明对比，两者都具有单一的具有统一连接六糖结构的硫酸软骨素链，增量六α1-3GalNAc(4-硫酸盐)β1-4GlcAβ1-3Gal(4-硫酸盐)β1-3Galβ1-4Xyl，含有4-O-硫酸基半乳糖残基。
Decorin is a small fibroblast proteoglycan consisting of a core protein and a single chondroitin/dermatan sulfate chain, The structure of the carbohydrate-protein linkage region of the recombinant decorin expressed in Chinese hamster ovary cells was investigated, The decorin was secreted in the culture medium and isolated by anion-exchange chromatography. The glycosaminoglycan chain was released from the decorin by beta-elimination using alkaline NaBH4, and then digested with chondroitinase ABC. These treatments resulted in a major and a few minor hexasaccharide alditols derived from the carbohydrate-protein linkage region, Their structures were analyzed by enzymatic digestion in conjunction with high-performance liquid chromatography, Two of these compounds have the conventional hexasaccharide core, Delta HexA alpha 1-3GalNAc beta 1-4GlcA beta 1-3GalNAc beta 1- -3Gal beta 1-4Xyl-ol. One is nonsulfated, and the other is monosulfated on C4 of the GalNAc residue, They represent 12% and 60% of the total linkage region, respectively, The other compound has the hexasaccharide alditol with an internal iduronic acid residue Delta HexA alpha 1-->3GalNAc(4-sulfate)beta 1-4IdoA alpha 1-3Gal beta 1-3Gal beta 1-4Xyl-ol, which was previously demonstrated in one of the five linkage hexasaccharide alditols isolated from dermatan sulfate proteoglycans of bovine aorta (Sugahara et al., J. Biol. Chem., 270, 7204-7212, 1995), The compound accounts for 11% of the total linkage region, These structural variations in the linkage hexasaccharide region of the decorin strikingly contrast to the uniformity demonstrated in the linkage hexasaccharide structure of human inter-alpha-trypsin inhibitor (Yamada ef al., Glycobiology, 5, 335-341, 1995) and urinary trypsin inhibitor (Yamada ef al., Eur, J. Biochem., 233, 687-693, 1995), both of which have a single chondroitin sulfate chain with a uniform linkage hexasaccharide structure, Delta HexA alpha 1-3GalNAc(4-sulfate)beta 1-4GlcA beta 1-3Gal(4-sulfate)beta 1-3Gal beta 1-4Xyl, containing a 4-O-sulfated Gal residue.