期刊
JOURNAL OF BIOLOGICAL CHEMISTRY
卷 276, 期 31, 页码 28806-28813出版社
AMER SOC BIOCHEMISTRY MOLECULAR BIOLOGY INC
DOI: 10.1074/jbc.M100204200
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资金
- NHLBI NIH HHS [5P01-HL41484, 5R01-HL58479] Funding Source: Medline
- NIGMS NIH HHS [R37-GM33063] Funding Source: Medline
To understand how 2-O-sulfation of uronic acid residues influences the biosynthesis of anticoagulant heparan sulfate, the cDNA encoding glucosaminyl 3-O-sulfotransferase-1 (3-OST-1) was introduced into wild-type Chinese hamster ovary cells and mutant pgsF-17 cells, which are defective in 2-O-sulfation. 3-OST-1-transduced cells gained the ability to bind to antithrombin. Structural analysis of the heparan sulfate chains showed that 3-OST-1 generates sequences containing GlcUA-GlcN(SO3)3(SO3) and GlcUA-GlcN(SO3)3(SO3)6(SO.) in both wild-type and mutant cells. In addition, IdoUA-GIcN(SO3)3(SO3) and IdoUA-GIcN(SO3)3(SO3)6(SO3) accumulate in the mutant chain. These disaccharides were also observed by tagging [6-H-3]GIcN-labeled pgsF-17 heparan sulfate in vitro with [S-35]PAPs and purified 3-OST-1. Heparan sulfate derived from the transduced mutant also had similar to2-fold higher affinity for antithrombin than heparan sulfate derived from the transduced wildtype cells, and it inactivated factor Xa more efficiently. This study demonstrates for the first time that (i) 3-O-sulfation by 3-OST-1 can occur independently of the 2-O-sulfation of uronic acids, (ii) 2-0-sulfation usually occurs before 3-O-sulfation, (iii) 2-0-sulfation blocks the action of 3-OST-1 at glucosamine residues located to the reducing side of IdoUA units, and (iv) that alternative antithrombin-binding structures can be made in the absence of 2-0-sulfation.
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