Bioengineering murine mastocytoma cells to produce anticoagulant heparin
| Autor: | Robert J. Linhardt, Payel Datta, Jeffrey D. Esko, Bo Yang, Jong Youn Baik, Trent R. Gemmill, Guoyun Li, Charles A. Glass, Susan T. Sharfstein, Leyla Gasimli |
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| Rok vydání: | 2013 |
| Předmět: |
Biochemistry & Molecular Biology
sulfotransferase Protein Engineering Medical and Health Sciences Biochemistry Dermatan sulfate Cell Line Mice chemistry.chemical_compound Glucosamine Cell Line Tumor Carbohydrate Conformation medicine Animals Tumor proteoglycan biology Heparin Chemistry mastocytoma Anticoagulants Mastocytoma Original Articles Hematology Transfection Heparan sulfate Biological Sciences medicine.disease Molecular biology Recombinant Proteins Proteoglycan Cell culture biology.protein heparan sulfate Sulfotransferases Biotechnology medicine.drug |
| Zdroj: | Glycobiology, vol 24, iss 3 |
| ISSN: | 1460-2423 0959-6658 |
| Popis: | Heparin (HP), an important anticoagulant polysaccharide, is produced in a complex biosynthetic pathway in connective tissue-type mast cells. Both the structure and size of HP are critical factors determining the anticoagulation activity. A murine mastocytoma (MST) cell line was used as a model system to gain insight into this pathway. As reported, MST cells produce a highly sulfated HP-like polysaccharide that lacks anticoagulant activity (Montgomery RI, Lidholt K, Flay NW, Liang J, Vertel B, Lindahl U, Esko JD. 1992. Stable heparin-producing cell lines derived from the Furth murine mastocytoma. Proc Natl Acad Sci USA 89:11327-11331). Here, we show that transfection of MST cells with a retroviral vector containing heparan sulfate 3-O-sulfotransferase-1 (Hs3st1) restores anticoagulant activity. The MST lines express N-acetylglucosamine N-deacetylase/N-sulfotransferase-1, uronosyl 2-O-sulfotransferase and glucosaminyl 6-O-sulfotransferase-1, which are sufficient to make the highly sulfated HP. Overexpression of Hs3st1 in MST-10H cells resulted in a change in the composition of heparan sulfate (HS)/HP and CS/dermatan sulfate (DS) glycosaminoglycans. The cell-associated HS/HP closely resembles HP with 3-O-sulfo group-containing glucosamine residues and shows anticoagulant activity. This study contributes toward a better understanding of the HP biosynthetic pathway with the goal of providing tools to better control the biosynthesis of HP chains with different structures and activities. |
| Databáze: | OpenAIRE |
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