Selective Effects of Sodium Chlorate Treatment on the Sulfation of Heparan Sulfate

• Published in: The Journal of biological chemistry Vol. 274; no. 51; pp. 36267 - 36273
• Main Authors: Safaiyan, F, Kolset, S O, Prydz, K, Gottfridsson, E, Lindahl, U, Salmivirta, M
• Format: Journal Article
• Language: English
• Published: United States American Society for Biochemistry and Molecular Biology 17.12.1999
• Subjects: Animals; Cell Line; Dogs; Heparitin Sulfate - chemistry; Hydrogen-Ion Concentration; Sodium Chloride - chemistry; Sodium Chloride - pharmacology; Sulfates
• ISSN: 0021-9258; 1083-351X
• DOI: 10.1074/jbc.274.51.36267

We have analyzed the effect of sodium chlorate treatment of Madin-Darby canine kidney cells on the structure of heparan sulfate (HS), to assess how the various sulfation reactions during HS biosynthesis are affected by decreased availability of the sulfate donor 3′-phosphoadenosine 5′-phosphosulfate. Metabolically [ 3 H]glucosamine-labeled HS was isolated from chlorate-treated and untreated Madin-Darby canine kidney cells and subjected to low pH nitrous acid cleavage. Saccharides representing (i) the N -sulfated domains, (ii) the domains of alternating N -acetylated and N -sulfated disaccharide units, and (iii) the N -acetylated domains were recovered and subjected to compositional disaccharide analysis. Upon treatment with 50 m m chlorate, overall O -sulfation of HS was inhibited by ∼70%, whereas N -sulfation remained essentially unchanged. Low chlorate concentrations (5 or 20 m m ) selectively reduced the 6- O -sulfation of HS, whereas treatment with 50 m m chlorate reduced both 2- O - and 6- O -sulfation. Analysis of saccharides representing the different domain types indicated that 6- O -sulfation was preferentially inhibited in the alternating domains. These data suggest that reduced 3′-phosphoadenosine 5′-phosphosulfate availability has distinct effects on the N - and O -sulfation of HS and that O -sulfation is affected in a domain-specific fashion.