Mouse Mastocytoma Cells Synthesize Undersulfated Heparin and Chondroitin Sulfate in the Presence of Brefeldin A

Uhlin‐Hansen, Lars; Kusche-Gullberg, Marion; Berg, Eli; Eriksson, Inger; Kjellén, Lena

doi:10.1074/jbc.272.6.3200

Cited by 17 publications

(17 citation statements)

References 34 publications

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“…NDST-1 was recently localized to the trans-Golgi network in NDST-1 transfected murine fibroblasts (38), such that at least some N-sulfation would occur as a late Golgi event. On the other hand, mastocytoma cells treated with Brefeldin A (which blocks transport from Golgi to the trans-Golgi network) produce heparin with N-, 2-O-and 6-O-sulfate groups (39), indicating that these sulfotransferase activities are also expressed proximal to the trans-Golgi network.…”

Section: Differential Effects Of Chlorate Treatment On 2-o-and 6-osulmentioning

confidence: 99%

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

Safaiyan¹,

Kolset²,

Prydz³

et al. 1999

Journal of Biological Chemistry

148

122

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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 mM chlorate, overall O-sulfation of HS was inhibited by ϳ70%, 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-Osulfation 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.

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Section: Differential Effects Of Chlorate Treatment On 2-o-and 6-osulmentioning

confidence: 99%

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

Safaiyan¹,

Kolset²,

Prydz³

et al. 1999

Journal of Biological Chemistry

148

122

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“…For example, cores that bear HS include glypicans, syndecans, perlecan, and agrin; cores that bear CS include members of the aggrecan and decorin families, among others. Some PGs, such as syndecan-1, commonly possess both HS and CS at different sites on the protein (6, 7), whereas others, such as serglycin, bear either CS or HS, depending upon the cell type in which they are expressed (8,9). Finally, some core proteins may be produced as a mixture of molecules possessing and lacking GAGs, or may lack GAGs entirely in some cell types.…”

Section: Proteoglycans (Pgs)mentioning

confidence: 99%

Mechanisms Underlying Preferential Assembly of Heparan Sulfate on Glypican-1

Chen¹,

Lander

2001

Journal of Biological Chemistry

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Glypicans are major cell surface heparan sulfate proteoglycans, the structures of which are characterized by the presence of a cysteine-rich globular domain, a short glycosaminoglycan (GAG) attachment region, and a glycosylphosphatidylinositol membrane anchor. Despite strong evolutionary conservation of the globular domains of glypicans, no function has yet been attributed to them. By using a novel quantitative approach for assessing proteoglycan glycosylation, we show here that removal of the globular domain from rat glypican-1 converts the proteoglycan from one that bears ϳ90% heparan sulfate (HS) to one that bears ϳ90% chondroitin sulfate. Mutational analysis shows that sequences at least 70 amino acids away from the glypican-1 GAG attachment site are required for preferential HS assembly, although more nearby sequences also play a role. The effects of the glypican-1 globular domain on HS assembly could also be demonstrated by fusing this domain to sequences representing the GAG attachment sites of other proteoglycans or, surprisingly, simply by expressing the isolated globular domain in cells and analyzing effects either on an exogenously expressed glypican-1 GAG attachment domain or on endogenous proteoglycans. Quantitative analysis of the effect of the globular domain on GAG addition to proteoglycan core proteins suggested that preferential HS assembly is achieved, at least in part, through the inhibition of chondroitin sulfate assembly. These data identify the glypican-1 globular domain as a structural motif that potently influences GAG class determination and suggest that an important role of glypican globular domains is to ensure a high level of HS substitution of these proteoglycans.

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“…Addition of brefeldin A to various cell cultures resulted in variable effects including shorter and undersulfated chondroitin/dermatan (36,37) and considerable loss of chondroitin sulfate-synthesing activity (22), suggesting considerable differences in specific Golgi subcompartments and between various specialized cells.…”

Section: Polysaccharide Formationmentioning

confidence: 99%

Biosynthesis of Chondroitin/Dermatan Sulfate

Silbert¹,

Sugumaran²

2002

IUBMB Life

292

234

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SummaryChondroitin sulfate and dermatan sulfate are synthesized as galactosaminoglycan polymers containing N-acetylgalactosmine alternating with glucuronic acid. The sugar residues are sulfated to varying degrees and positions depending upon the tissue sources and varying conditions of formation. Epimerization of any of the glucuronic acid residues to iduronic acid at the polymer level constitutes the formation of dermatan sulfate. Chondroitin/dermatan glycosaminoglycans are covalently attached by a common tetrasaccharide sequence to the serine residues of core proteins while they are adherent to the inner surface of endoplasmic reticulum/Golgi vesicles. Addition of the first sugar residue, xylose, to core proteins begins in the endoplasmic reticulum, followed by the addition of two galactose residues by two distinct glycosyl transferases in the early cis/medial regions of the Golgi. The linkage tetrasaccharide is completed in the medial/trans Golgi by the addition of the first glucuronic acid residue, followed by transfer of N-acetylgalactosamine to initiate the formation of a galactosaminoglycan rather than a glucosaminoglycan. This specific N-acetylgalactosaminyl transferase is different from the chondroitin synthase involved in generation of the repeating disaccharide units to form the chondroitin polymer. Sulfation of the chondroitin polymer by specific sulfotransferases occurs as the polymer is being formed. All the enzymes in the pathway for synthesis have been cloned, with the exception of the glucuronyl to iduronyl epimerase involved in the formation of dermatan residues. IUBMB Life, 54: 177-186, 2002

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Mouse Mastocytoma Cells Synthesize Undersulfated Heparin and Chondroitin Sulfate in the Presence of Brefeldin A

Abstract: In order to study the subcellular localization and organization of the enzymes involved in the glycosylation of the hybrid proteoglycan serglycin, mouse mastocytoma cells were metabolically labeled with [

Cited by 17 publications

References 34 publications

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

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

Mechanisms Underlying Preferential Assembly of Heparan Sulfate on Glypican-1

Biosynthesis of Chondroitin/Dermatan Sulfate

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