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Structure and Biosynthesis of Carbohydrate-Protein Linkage Region of Dermatan Sulfate and Chondroitin Sulfate

硫酸皮肤素和硫酸软骨素碳水化合物-蛋白质连接区的结构和生物合成

基本信息

批准号：
06808061
负责人：
SUGAHARA Kazuyuki
金额：
$ 1.22万
依托单位：
Kobe Pharmaceutical University
依托单位国家：
日本
项目类别：
Grant-in-Aid for General Scientific Research (C)
财政年份：
1994
资助国家：
日本
起止时间：
1994 至 1995
项目状态：
已结题

项目摘要

The fine structure and the biosynthetic mechanisms of sulfated glycosamino-glycans are not fully understood. In this study, we carried out structural studies on the carbohydrate-protein linkage region of a dermatan sulfate preparation and various chondroitin sulfate preparations, and also studied putative biosynthetic enzymes.In the structural studies, we isolated five hexasaccharide alditols derived from the linkage region of bovine aorta dermatan sulfate chains after chondoritinase ABC digestion and beta-elimination reaction. The structures of the three of them were identical to those isolated previously from chondroitin 4-sulfate of whale cartilage. In contrast, the other two contained hitherto unreported structures including iduronic acid (IdoA) : DELTAHexAalpha1-3GalNAc (4-sulfate) beta1-4IdoAalpha1-3Gal (4-sulfate) beta1-3Galbeta1-4Xyl-ol and DELTAHexAalpha1-3GalNAc (4-sulfate) beta1-4IdoAalpha1-3Galbeta1-3Galbeta1-4Xyl-ol.We also isolated the following hexasaccharide as a sole linkage component from chondroitin 4-sulfate of inter alpha-trypsin inhibitor in human plasma and a trypsin inhibitor in human urine : DELTAHexAalpha1-3GalNAc (4-omicron-sulfate) beta1-4GlcAbeta1-3Gal (4-omicron-sulfate) beta1-3Galbeta1-4Xyl-ol. The results are taken to indicate the importance of the sulfated Gal structure in the biosynthesis of the linkage region.In the biosynthetic studies, we discovered a hitherto unreported alpha-GalNAc transferase activity in fetal bovine serum. It catalyzed a transfer of alpha-GlaNAc residue to GlcAbeta1-3Galbeta1-3Galbeta1-4Xylbeta1-omicron-Ser, but not to GlcAbeta1-3Gal (4-omicron-sulfate) beta1-3Galbeta1-omicron-Ser. It remains to be investigated whether or not this enzyme is in involved indeed in the biosynthesis of chondroitin sulfate. In addition, we detected in fetal bovine serum a beta-GalNAc transferase activity that is involved in the synthesis of the repeating disaccharide region of chondroitin sulfate.

巯基糖胺聚糖的精细结构和生物合成机理尚不完全清楚。在本研究中，我们对硫酸皮炎制剂和各种硫酸软骨素制剂的糖-蛋白连锁区进行了结构研究，并对可能的生物合成酶进行了研究。在结构研究中，我们从牛主动脉皮聚糖硫酸链的连锁区分离出5个经过软骨素酶ABC消化和β消除反应的六糖醛醇。其中三个的结构与先前从鲸鱼软骨中分离出的硫酸软骨素4的结构相同。相比之下，其他两个含有迄今未报道的结构，包括伊杜醛酸（IdoA）： DELTAHexAalpha1-3GalNAc（4-硫酸盐）beta1-4IdoAalpha1-3Gal（4-硫酸盐）beta1-3Galbeta1-4Xyl-ol和DELTAHexAalpha1-3GalNAc（4-硫酸盐）beta1-4IdoAalpha1-3Galbeta1-3Galbeta1-4Xyl-ol。我们还从人血浆中α -胰蛋白酶抑制剂和人尿中胰蛋白酶抑制剂的4-硫酸软骨素中分离出以下六糖作为唯一的连锁成分：DELTAHexAalpha1-3GalNAc (4- omicon -sulfate) β 1- 4glcabeta1 - 3gal (4- omicon -sulfate) β 1- 3galbeta1 - 4xyl -ol。这些结果表明了硫酸化Gal结构在连接区生物合成中的重要性。在生物合成研究中，我们在胎牛血清中发现了迄今未报道的α - galnac转移酶活性。它催化了α - glanac残基向glcabeta1 - 3galbeta1 - 3galbeta1 - 4xylβ - omicon - ser的转移，但没有向GlcAbeta1-3Gal (4- omicon -sulfate) β - 1- 3galbeta1 - omicon - ser的转移。这种酶是否确实参与硫酸软骨素的生物合成还有待研究。此外，我们在胎牛血清中检测到β - galnac转移酶活性，该酶参与硫酸软骨素重复双糖区域的合成。