Characterization of a cell mutant specifically defective in the synthesis of anticoagulantly active heparan sulfate
- PMID: 7929178
Characterization of a cell mutant specifically defective in the synthesis of anticoagulantly active heparan sulfate
Abstract
We have investigated the characteristics of clonal L cell mutant VI-7 that has previously been demonstrated to exhibit reduced levels of cell surface proteoglycans containing heparan sulfate (HS) with regions of defined monosaccharide sequence that interact with antithrombin (HSact) (de Agostini, A. L., Lau, H. K., Leone, C., Youssoufian, H., and Rosenberg, R. D. (1990) Proc. Natl. Acad. Sci. U.S.A. 87, 9784-9788). Pulse labeling revealed that the synthesis of HSact in mutant cells, as compared to wild-type cells, was reduced by 5-7-fold, which is identical to the previously observed reduction in cell surface-bound antithrombin. This alteration is independent of growth state since production of HSact by mutant cells, as compared to wild-type cells, is decreased to a similar extent in exponentially growing and post-confluent cultures. The synthetic defect was specific for HSact since production of total HS and chondroitin sulfate by mutant cells, as compared to wild-type cells, was identical in magnitude. The synthetic abnormality is not due to an alteration in core protein since complementation was not observed when mutant cells were stably transfected with the epitope-tagged ryudocan cDNA which can initiate HSact production. Structural analyses revealed that HS from mutant cells, as compared to wild-type cells, exhibited normal molecular weight, extent and distribution of sulfate, and disaccharide composition, which indicate that the mutation did not affect HS biosynthetic enzymes. Together, the data suggest that mutant VI-7 is defective in a regulatory component which directly or indirectly coordinates HS biosynthetic enzymes to specifically generate the defined monosaccharide sequence of HSact.
Similar articles
-
Pathway-specific regulation of the synthesis of anticoagulantly active heparan sulfate.J Biol Chem. 1994 Oct 7;269(40):24941-52. J Biol Chem. 1994. PMID: 7929177
-
Cell-free synthesis of anticoagulant heparan sulfate reveals a limiting converting activity that modifies an excess precursor pool.J Biol Chem. 1996 Oct 25;271(43):27063-71. J Biol Chem. 1996. PMID: 8900197
-
Isolation and characterization of ryudocan and syndecan heparan sulfate proteoglycans, core proteins, and cDNAs from a rat endothelial cell line.Haemostasis. 1993 Mar;23 Suppl 1:161-76. doi: 10.1159/000216925. Haemostasis. 1993. PMID: 8495865
-
The retinoic acid and cAMP-dependent up-regulation of 3-O-sulfotransferase-1 leads to a dramatic augmentation of anticoagulantly active heparan sulfate biosynthesis in F9 embryonal carcinoma cells.J Biol Chem. 1998 Oct 23;273(43):27998-8003. doi: 10.1074/jbc.273.43.27998. J Biol Chem. 1998. PMID: 9774414
-
Molecular biology of ryudocan, an endothelial heparan sulfate proteoglycan.Semin Thromb Hemost. 2000;26(1):67-73. doi: 10.1055/s-2000-9806. Semin Thromb Hemost. 2000. PMID: 10805285 Review.
Cited by
-
Substrate specificity of 6-O-endosulfatase (Sulf-2) and its implications in synthesizing anticoagulant heparan sulfate.Glycobiology. 2012 Oct;22(10):1353-62. doi: 10.1093/glycob/cws092. Epub 2012 Jun 12. Glycobiology. 2012. PMID: 22692045 Free PMC article.
-
Discovery of a heparan sulfate 3-O-sulfation specific peeling reaction.Anal Chem. 2015 Jan 6;87(1):592-600. doi: 10.1021/ac503248k. Epub 2014 Dec 22. Anal Chem. 2015. PMID: 25486437 Free PMC article.
-
Differential roles for 3-OSTs in the regulation of cilia length and motility.Development. 2013 Sep;140(18):3892-902. doi: 10.1242/dev.096388. Epub 2013 Aug 14. Development. 2013. PMID: 23946439 Free PMC article.
-
Normal levels of anticoagulant heparan sulfate are not essential for normal hemostasis.J Clin Invest. 2003 Apr;111(7):989-99. doi: 10.1172/JCI15809. J Clin Invest. 2003. PMID: 12671048 Free PMC article.
-
Synthesis of 3-O-Sulfated Disaccharide and Tetrasaccharide Standards for Compositional Analysis of Heparan Sulfate.Biochemistry. 2020 Sep 1;59(34):3186-3192. doi: 10.1021/acs.biochem.9b00838. Epub 2019 Oct 23. Biochemistry. 2020. PMID: 31608625 Free PMC article.
Publication types
MeSH terms
Substances
Grants and funding
LinkOut - more resources
Full Text Sources
Other Literature Sources
Medical