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Bioengineering murine mastocytoma cells to produce anticoagulant heparin.
MedLine Citation:
PMID:  24326668     Owner:  NLM     Status:  Publisher    
Abstract/OtherAbstract:
Heparin, an important anticoagulant polysaccharide, is produced in a complex biosynthetic pathway in connective tissue-type mast cells. Both the structure and size of heparin are critical factors determining the anticoagulation activity. A murine mastocytoma (MST) cell line was used as a model system to gain insight into the details of this pathway. As reported previously, MST cells produce a highly sulfated heparin-like polysaccharide that lacks anticoagulant activity (Montgomery et al. 1992. Proc Natl Acad Sci USA 89:11327). 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 heparin. Overexpression of Hs3st1 in MST-10H cells resulted in a change in the composition of HS/HP and CS/DS glycosaminoglycans. The cell associated HS/HP closely resembles heparin with 3-O-sulfo group containing glucosamine residues and shows anticoagulant activity. This study contributes towards 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.
Authors:
Leyla Gasimli; Charles A Glass; Payel Datta; Bo Yang; Guoyun Li; Trent R Gemmill; Jong Youn Baik; Susan T Sharfstein; Jeffrey D Esko; Robert J Linhardt
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Publication Detail:
Type:  JOURNAL ARTICLE     Date:  2013-12-9
Journal Detail:
Title:  Glycobiology     Volume:  -     ISSN:  1460-2423     ISO Abbreviation:  Glycobiology     Publication Date:  2013 Dec 
Date Detail:
Created Date:  2013-12-11     Completed Date:  -     Revised Date:  -    
Medline Journal Info:
Nlm Unique ID:  9104124     Medline TA:  Glycobiology     Country:  -    
Other Details:
Languages:  ENG     Pagination:  -     Citation Subset:  -    
Affiliation:
Department of Biology and Center for Biotechnology and Interdisciplinary Studies, Rensselaer Polytechnic Institute, 110 8th Avenue , Troy NY 12180, USA.
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