| Project/Area Number |
06808061
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| Research Category |
Grant-in-Aid for General Scientific Research (C)
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| Allocation Type | Single-year Grants |
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| Research Field |
Structural biochemistry
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| Research Institution | Kobe Pharmaceutical University |
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Principal Investigator |
SUGAHARA Kazuyuki Kobe Pharmaceutical Univ.Professor Dept.of Biochemistry, 薬学部・生化学, 教授 (60154449)
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| Co-Investigator(Kenkyū-buntansha) |
YAMADA Shuhei Kobe Pharmaceutical Univ.Instructor Dept.of Biochemistry, 薬学部・生化学, 助手 (70240017)
KITAGAWA Hiroshi Kobe Pharmaceutical Univ.Assistant Professor Dept.of Biochemistry, 薬学部・生化学, 講師 (40221915)
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| Project Period (FY) |
1994 – 1995
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| Project Status |
Completed (Fiscal Year 1995)
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| Budget Amount *help |
¥1,900,000 (Direct Cost: ¥1,900,000)
Fiscal Year 1995: ¥1,000,000 (Direct Cost: ¥1,000,000)
Fiscal Year 1994: ¥900,000 (Direct Cost: ¥900,000)
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| Keywords | Glycosaminoglycan / Dermatan Sulfate / Chondroitin Sulfate / Sulfation / Sulfated Oligosaccharides / ウリナリートリプシンインヒビタ / GalNAc転移酵素 / インターα-トリプシンインヒビター / ウリマリ-トリプシンインヒビター / グリコサミノグリカン / ESI / MS / NMR |
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| Research Abstract |
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.
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