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

doi:10.1074/jbc.274.51.36267

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

From the ^‡Department of Medical Biochemistry and Microbiology, Uppsala University, Biomedical Center, P. O. Box 582, S-75123 Uppsala, Sweden and the ^§Institute for Nutrition Research and ^¶Department of Biochemistry, University of Oslo, N-0316 Oslo, Norway

Abstract

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 [³H]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 alternatingN-acetylated and N-sulfated disaccharide units, and (iii) the N-acetylated domains were recovered and subjected to compositional disaccharide analysis. Upon treatment with 50 mm chlorate, overall O-sulfation of HS was inhibited by ∼70%, whereas N-sulfation remained essentially unchanged. Low chlorate concentrations (5 or 20 mm) selectively reduced the 6-O-sulfation of HS, whereas treatment with 50 mm 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 thatO-sulfation is affected in a domain-specific fashion.

Footnotes

↵* This work was supported by Grants K96–03P, K99–03X, and 2309 from the Swedish Medical Research Council; Grant 3919-B97 from the Swedish Society for Cancer Research; The Medical Faculty of Uppsala University; Polysackaridforskning AB (Uppsala, Sweden); and The Norwegian Cancer Society.The costs of publication of this article were defrayed in part by the payment of page charges. The article must therefore be hereby marked “advertisement” in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.
↵‖ To whom correspondence should be addressed. Turku Centre for Biotechnology Tykistökatu 6, BioCity FIN-20520 Turku, Finland. Tel.: 358-2-3338047; Fax: 358-2-3338000; E-mail: markku.salmivirta@btk. utu.fi.
↵2 U. Lindahl, unpublished data.
Abbreviations:

HS

heparan sulfate

aMan_R

2,5-anhydromannitol

GlcUA

d-glucuronic acid

GlcN

glucosamine

HexA

hexuronic acid

HPLC

high pressure liquid chromatography

IdoA

l-iduronic acid

MDCK

Madin-Darby canine kidney

NA

N-acetylated

NDST

glucosaminyl N-deacetylase/N-sulfotransferase

NS

N-sulfated

PAPS

3′-phosphoadenosine 5′-phosphosulfate
- Received June 25, 1999.
- Revision received September 9, 1999.
The American Society for Biochemistry and Molecular Biology, Inc.