title: NDST2 (N-Deacetylase/N-Sulfotransferase-2) Enzyme Regulates Heparan Sulfate Chain Length.
abstract: Analysis of heparan sulfate synthesized by HEK 293 cells overexpressing murine NDST1 and/or NDST2 demonstrated that the amount of heparan sulfate was increased in NDST2- but not in NDST1-overexpressing cells. Altered transcript expression of genes encoding other biosynthetic enzymes or proteoglycan core proteins could not account for the observed changes. However, the role of NDST2 in regulating the amount of heparan sulfate synthesized was confirmed by analyzing heparan sulfate content in tissues isolated from Ndst2(-/-) mice, which contained reduced levels of the polysaccharide. Detailed disaccharide composition analysis showed no major structural difference between heparan sulfate from control and Ndst2(-/-) tissues, with the exception of heparan sulfate from spleen where the relative amount of trisulfated disaccharides was lowered in the absence of NDST2. In vivo transcript expression levels of the heparan sulfate-polymerizing enzymes Ext1 and Ext2 were also largely unaffected by NDST2 levels, pointing to a mode of regulation other than increased gene transcription. Size estimation of heparan sulfate polysaccharide chains indicated that increased chain lengths in NDST2-overexpressing cells alone could explain the increased heparan sulfate content. A model is discussed where NDST2-specific substrate modification stimulates elongation resulting in increased heparan sulfate chain length.
authors.affiliation: From the Department of Medical Biochemistry and Microbiology, Science for Life Laboratory, Uppsala University, SE-75123 Uppsala, Sweden and.
authors.forename: Audrey
authors.initials: A
authors.lastname: Deligny
authors.affiliation: From the Department of Medical Biochemistry and Microbiology, Science for Life Laboratory, Uppsala University, SE-75123 Uppsala, Sweden and.
authors.forename: Tabea
authors.initials: T
authors.lastname: Dierker
authors.affiliation: From the Department of Medical Biochemistry and Microbiology, Science for Life Laboratory, Uppsala University, SE-75123 Uppsala, Sweden and.
authors.forename: Anders
authors.initials: A
authors.lastname: Dagälv
authors.affiliation: From the Department of Medical Biochemistry and Microbiology, Science for Life Laboratory, Uppsala University, SE-75123 Uppsala, Sweden and.
authors.forename: Anders
authors.initials: A
authors.lastname: Lundequist
authors.affiliation: From the Department of Medical Biochemistry and Microbiology, Science for Life Laboratory, Uppsala University, SE-75123 Uppsala, Sweden and.
authors.forename: Inger
authors.initials: I
authors.lastname: Eriksson
authors.affiliation: the Complex Carbohydrate Research Center, University of Georgia, Athens, Georgia 30602.
authors.forename: Alison V
authors.initials: AV
authors.lastname: Nairn
authors.affiliation: the Complex Carbohydrate Research Center, University of Georgia, Athens, Georgia 30602.
authors.forename: Kelley W
authors.initials: KW
authors.lastname: Moremen
authors.affiliation: From the Department of Medical Biochemistry and Microbiology, Science for Life Laboratory, Uppsala University, SE-75123 Uppsala, Sweden and.
authors.forename: Catherine L R
authors.initials: CLR
authors.lastname: Merry
authors.affiliation: From the Department of Medical Biochemistry and Microbiology, Science for Life Laboratory, Uppsala University, SE-75123 Uppsala, Sweden and lena.kjellen@imbim.uu.se.
authors.forename: Lena
authors.initials: L
authors.lastname: Kjellén
mesh_terms: D000581:Amidohydrolases / Q000096:biosynthesis* / Q000235:genetics; D000818:Animals; D015971:Gene Expression Regulation, Enzymologic / Q000502:physiology*; D057809:HEK293 Cells; D006497:Heparan Sulfate / Q000096:biosynthesis* / Q000235:genetics; D006801:Humans; D051379:Mice; D018345:Mice, Knockout; D008954:Models, Biological; D017351:N-Acetylglucosaminyltransferases / Q000096:biosynthesis / Q000235:genetics; D015238:Sulfotransferases / Q000096:biosynthesis* / Q000235:genetics; D014158:Transcription, Genetic / Q000502:physiology*; D000099065:Exostosin 2; D000099058:Exostosin 1
chemical_list: D000581:Amidohydrolases; D006497:Heparan Sulfate; D017351:N-Acetylglucosaminyltransferases; D015238:Sulfotransferases; D000099065:Exostosin 2; D000099058:Exostosin 1; C491855:Ndst2 protein, mouse
keywords: Golgi; heparan sulfate; heparin; proteoglycan synthesis; sulfotransferase