Journal article
Biosynthesis of 3-O-sulfated heparan sulfate: unique substrate specificity of heparan sulfate 3-O-sulfotransferase isoform 5
Glycobiology (Oxford), Vol.13(11), pp.785-794
11/2003
DOI: 10.1093/glycob/cwg101
PMID: 12907690
Abstract
Heparan sulfate 3-O-sulfotransferase transfers sulfate to the 3-OH position of a glucosamine to generate 3-O-sulfated heparan sulfate (HS), which is a rare component in HS from natural sources. We previously reported that 3-O- sulfotransferase isoform 5 (3-OST-5) generates both an antithrombin-binding site to exhibit anticoagulant activity and a binding site for herpes simplex virus 1 glycoprotein D to serve as an entry receptor for herpes simplex virus. In this study, we characterize the substrate specificity of 3-OST-5 using the purified enzyme. The enzyme was expressed in insect cells using the baculovirus expression approach and was purified by using heparin-Sepharose and 3′,5′-ADP- agarose chromatographies. As expected, the purified enzyme generates both an antithrombin binding site and a glycoprotein D binding site. We isolated IdoUA-AnMan3S and IdoUA-AnMan3S6S from nitrous acid–degraded 3-OST-5-modified HS (pH 1.5), suggesting that 3-OST-5 enzyme sulfates the glucosamine residue that is linked to an iduronic acid residue at the nonreducing end. We also isolated a disaccharide with a structure of ΔUA2S-GlcNS3S and a tetrasaccharide with a structure of ΔUA2S-GlcNS-IdoUA2S-GlcNH23S6S from heparin lyases–digested 3-OST-5-modified HS. Our results suggest that 3-OST-5 enzyme sulfates both N-sulfated glucosamine and N-unsubstituted glucosamine residues. Taken together, the results indicate that 3-OST-5 has broader substrate specificity than those of 3-OST-1 and 3-OST-3. The unique substrate specificity of 3-OST-5 serves as an additional tool to study the mechanism for the biosynthesis of biologically active HS.
Details
- Title: Subtitle
- Biosynthesis of 3-O-sulfated heparan sulfate: unique substrate specificity of heparan sulfate 3-O-sulfotransferase isoform 5
- Creators
- Jinghua Chen - University of North Carolina at Chapel HillMichael B. DuncanKevin Carrick - Proteomic Core Facility, Department of Biochemistry, University of North Carolina, Chapel Hill, NC 27599R. Marshall Pope - Proteomic Core Facility, Department of Biochemistry, University of North Carolina, Chapel Hill, NC 27599Jian Liu
- Resource Type
- Journal article
- Publication Details
- Glycobiology (Oxford), Vol.13(11), pp.785-794
- Publisher
- Oxford University Press
- DOI
- 10.1093/glycob/cwg101
- PMID
- 12907690
- ISSN
- 0959-6658
- eISSN
- 1460-2423
- Language
- English
- Date published
- 11/2003
- Academic Unit
- Medicine Administration
- Record Identifier
- 9984627346102771
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