title: Heparan sulfate structure: methods to study N-sulfation and NDST action.
abstract: Heparan sulfate proteoglycans are important modulators of cellular processes where the negatively charged polysaccharide chains interact with target proteins. The sulfation pattern of the heparan sulfate chains will determine the proteins that will bind and the affinity of the interactions. The N-deacetylase/N-sulfotransferase (NDST) enzymes are of key importance during heparan sulfate biosynthesis when the sulfation pattern is determined. In this chapter, metabolic labeling of heparan sulfate with [(35)S]sulfate or [(3)H]glucosamine in cell cultures is described, in addition to characterization of polysaccharide chain length and degree of N-sulfation. Methods to measure NDST enzyme activity are also presented.
authors.affiliation: Department of Medical Biochemistry and Microbiology, Science for Life Laboratory, Uppsala University, 256, 751 23, Uppsala, Sweden.
authors.forename: Anders
authors.initials: A
authors.lastname: Dagälv
authors.forename: Anders
authors.initials: A
authors.lastname: Lundequist
authors.forename: Beata
authors.initials: B
authors.lastname: Filipek-Górniok
authors.forename: Tabea
authors.initials: T
authors.lastname: Dierker
authors.forename: Inger
authors.initials: I
authors.lastname: Eriksson
authors.forename: Lena
authors.initials: L
authors.lastname: Kjellén
mesh_terms: D002460:Cell Line; D002850:Chromatography, Gel; D004797:Enzyme-Linked Immunosorbent Assay; D006497:Heparan Sulfate / Q000096:biosynthesis / Q000737:chemistry*; D006801:Humans; D007553:Isotope Labeling / Q000379:methods*; D013431:Sulfates / Q000378:metabolism*; D015238:Sulfotransferases / Q000378:metabolism*; D013462:Sulfur Radioisotopes; D014316:Tritium
chemical_list: D006497:Heparan Sulfate; D013431:Sulfates; D015238:Sulfotransferases; D013462:Sulfur Radioisotopes; D014316:Tritium