The biosynthesis of anticoagulant heparan sulfate by the heparan sulfate 3-O-sulfotransferase isoform 5

Michael B. Duncan; Jinghua Chen; Jeffrey P. Krise; Jian Liu

doi:10.1016/j.bbagen.2003.12.010

The biosynthesis of anticoagulant heparan sulfate by the heparan sulfate 3-O-sulfotransferase isoform 5

Michael B. Duncan, Jinghua Chen, Jeffrey P. Krise, Jian Liu

Research output: Contribution to journal › Article › peer-review

34 Scopus citations

Abstract

The role of heparan sulfate (HS) in regulating blood coagulation has a wide range of clinical implications. In this study, we investigated the role of 3-O-sulfotransferase isoform 5 (3-OST-5) in generating anticoagulant HS in vivo. A Chinese hamster ovary cell line (3OST5/CHO) stably expressing 3-OST-5 was generated. The expression of 3-OST-5 in 3OST5/CHO cells was confirmed by Northern blot analysis, RT-PCR, and the disaccharide analyses of the HS from the cells. We also determined the effects of the HS from 3OST5/CHO on antithrombin-mediated inhibition of factor Xa. Fluorescently labeled antithrombin bound to the surface of 3OST5/CHO cells, suggesting that the antithrombin-binding HS is indeed present on the cell surface. Our results demonstrate that the 3-OST-5 gene is capable of synthesizing anticoagulant HS in CHO cells and has the potential to contribute to the biosynthesis of anticoagulant HS in humans.

Original language	English (US)
Pages (from-to)	34-43
Number of pages	10
Journal	Biochimica et Biophysica Acta - General Subjects
Volume	1671
Issue number	1-3
DOIs	https://doi.org/10.1016/j.bbagen.2003.12.010
State	Published - Mar 17 2004
Externally published	Yes

Keywords

3OST
3OST5/CHO
A CHO cell that stably expresses 3-OST-5
AT
Anticoagulation
Antithrombin
Blood coagulation
CHO
Chinese hamster ovary
HS
Heparan sulfate
Heparan sulfate 3-O-sulfotransferase
Heparan sulfate sulfotransferase

ASJC Scopus subject areas

Biophysics
Biochemistry
Molecular Biology

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10.1016/j.bbagen.2003.12.010

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title = "The biosynthesis of anticoagulant heparan sulfate by the heparan sulfate 3-O-sulfotransferase isoform 5",

abstract = "The role of heparan sulfate (HS) in regulating blood coagulation has a wide range of clinical implications. In this study, we investigated the role of 3-O-sulfotransferase isoform 5 (3-OST-5) in generating anticoagulant HS in vivo. A Chinese hamster ovary cell line (3OST5/CHO) stably expressing 3-OST-5 was generated. The expression of 3-OST-5 in 3OST5/CHO cells was confirmed by Northern blot analysis, RT-PCR, and the disaccharide analyses of the HS from the cells. We also determined the effects of the HS from 3OST5/CHO on antithrombin-mediated inhibition of factor Xa. Fluorescently labeled antithrombin bound to the surface of 3OST5/CHO cells, suggesting that the antithrombin-binding HS is indeed present on the cell surface. Our results demonstrate that the 3-OST-5 gene is capable of synthesizing anticoagulant HS in CHO cells and has the potential to contribute to the biosynthesis of anticoagulant HS in humans.",

keywords = "3OST, 3OST5/CHO, A CHO cell that stably expresses 3-OST-5, AT, Anticoagulation, Antithrombin, Blood coagulation, CHO, Chinese hamster ovary, HS, Heparan sulfate, Heparan sulfate 3-O-sulfotransferase, Heparan sulfate sulfotransferase",

author = "Duncan, {Michael B.} and Jinghua Chen and Krise, {Jeffrey P.} and Jian Liu",

note = "Funding Information: We acknowledge Mr. Ronald Copeland and Ms. Suzanne Thorp (University of North Carolina) for technical assistance and helpful discussion. The authors also thank Dr. Lijuan Zhang (Washington University, St. Louis) for helping us in preparing fluorescently labeled AT. This work was supported by an NIH grant (AI50050, to J.L.) and grant-in-aid (0355800U, to J.L.) from American Heart Association Mid-Atlantic Affiliate. M.D. is a recipient of Predoctoral Fellowship (2001-17095) from The David and Lucile Packard Foundation.",

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doi = "10.1016/j.bbagen.2003.12.010",

language = "English (US)",

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T1 - The biosynthesis of anticoagulant heparan sulfate by the heparan sulfate 3-O-sulfotransferase isoform 5

AU - Duncan, Michael B.

AU - Chen, Jinghua

AU - Krise, Jeffrey P.

AU - Liu, Jian

N1 - Funding Information: We acknowledge Mr. Ronald Copeland and Ms. Suzanne Thorp (University of North Carolina) for technical assistance and helpful discussion. The authors also thank Dr. Lijuan Zhang (Washington University, St. Louis) for helping us in preparing fluorescently labeled AT. This work was supported by an NIH grant (AI50050, to J.L.) and grant-in-aid (0355800U, to J.L.) from American Heart Association Mid-Atlantic Affiliate. M.D. is a recipient of Predoctoral Fellowship (2001-17095) from The David and Lucile Packard Foundation.

PY - 2004/3/17

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N2 - The role of heparan sulfate (HS) in regulating blood coagulation has a wide range of clinical implications. In this study, we investigated the role of 3-O-sulfotransferase isoform 5 (3-OST-5) in generating anticoagulant HS in vivo. A Chinese hamster ovary cell line (3OST5/CHO) stably expressing 3-OST-5 was generated. The expression of 3-OST-5 in 3OST5/CHO cells was confirmed by Northern blot analysis, RT-PCR, and the disaccharide analyses of the HS from the cells. We also determined the effects of the HS from 3OST5/CHO on antithrombin-mediated inhibition of factor Xa. Fluorescently labeled antithrombin bound to the surface of 3OST5/CHO cells, suggesting that the antithrombin-binding HS is indeed present on the cell surface. Our results demonstrate that the 3-OST-5 gene is capable of synthesizing anticoagulant HS in CHO cells and has the potential to contribute to the biosynthesis of anticoagulant HS in humans.

AB - The role of heparan sulfate (HS) in regulating blood coagulation has a wide range of clinical implications. In this study, we investigated the role of 3-O-sulfotransferase isoform 5 (3-OST-5) in generating anticoagulant HS in vivo. A Chinese hamster ovary cell line (3OST5/CHO) stably expressing 3-OST-5 was generated. The expression of 3-OST-5 in 3OST5/CHO cells was confirmed by Northern blot analysis, RT-PCR, and the disaccharide analyses of the HS from the cells. We also determined the effects of the HS from 3OST5/CHO on antithrombin-mediated inhibition of factor Xa. Fluorescently labeled antithrombin bound to the surface of 3OST5/CHO cells, suggesting that the antithrombin-binding HS is indeed present on the cell surface. Our results demonstrate that the 3-OST-5 gene is capable of synthesizing anticoagulant HS in CHO cells and has the potential to contribute to the biosynthesis of anticoagulant HS in humans.

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KW - Anticoagulation

KW - Antithrombin

KW - Blood coagulation

KW - CHO

KW - Chinese hamster ovary

KW - HS

KW - Heparan sulfate

KW - Heparan sulfate 3-O-sulfotransferase

KW - Heparan sulfate sulfotransferase

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IS - 1-3

ER -

The biosynthesis of anticoagulant heparan sulfate by the heparan sulfate 3-O-sulfotransferase isoform 5

Abstract

Keywords

ASJC Scopus subject areas

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