Steroid sulfatase, arylsulfatases A and B, galactose-6-sulfatase, and iduronate sulfatase in mammary cells and effects of sulfated and non-sulfated estrogens on sulfatase activity

Journal of Biological Chemistry

Kotlo

Shukla

et al. 2008

Self Cite

The sulfatase enzymes, N-acetylgalactosamine-4-sulfatase (arylsulfatase B (ASB)) and galactose-6-sulfatase (GALNS) hydrolyze sulfate groups of CS. Deficiencies of ASB and GALNS are associated with the mucopolysaccharidoses. To determine if expression of ASB and GALNS impacts on glycosaminoglycans (GAGs) and proteoglycans beyond their association with the mucopolysaccharidoses, we modified the expression of ASB and GALNS by overexpression and by silencing with small interference RNA in MCF-7 cells. Content of total sulfated GAG (sGAG), chondroitin 4-sulfate (C4S), and total chondroitin sulfates (CSs) was measured following immunoprecipitation with C4S and CS antibodies and treatment with chondroitinase ABC. Following silencing of ASB or GALNS, total sGAG, C4S, and CS increased significantly. Following overexpression of ASB or GALNS, total sGAG, C4S, and CS declined significantly. Measurements following chondroitinase ABC treatment of the cell lysates demonstrated no change in the content of the other sGAG, including heparin, heparan sulfate, dermatan sulfate, and keratan sulfate. Following overexpression of ASB and immunoprecipitation with C4S antibody, virtually no sGAG was detectable. Total sGAG content increased to 23.39 (؎1.06) g/mg of protein from baseline of 12.47 (؎0.68) g/mg of protein following ASB silencing. mRNA expression of core proteins of the CS-containing proteoglycans, syndecan-1 and decorin, was significantly up-regulated following overexpression of ASB and GALNS. Soluble syndecan-1 protein increased following increases in ASB and GALNS and reduced following silencing, inversely to changes in CS. These findings demonstrate that modification of expression of the lysosomal sulfatases ASB and GALNS regulates the content of CSs. N-Acetylgalactosamine-4-sulfatase (arylsulfatase B, ASB)2 and galactose-6-sulfatase (GALNS) are two important lysosomal enzymes that hydrolyze sulfate groups from the glycosaminoglycans (GAGs) chondroitin sulfate (CS) and dermatan sulfate. Deficiencies of ASB and GALNS activity are associated with the congenital mucopolysaccharidoses (MPS) MaroteauxLamy syndrome (MPS VI) and Morquio syndrome (MPS IVA), respectively. In these lysosomal storage diseases, there is accumulation of highly sulfated, unmetabolizable glycosaminoglycans and proteoglycans. The impact of sulfatase activity on metabolism of GAGs and the proteoglycans (PGs) with which they are associated has not been widely explored, beyond the congenital storage diseases.Recently, we reported that ASB activity is reduced in uncorrected cystic fibrosis cells, consistent with the accumulation of highly sulfated GAG in pulmonary secretions in cystic fibrosis patients. Following correction of cystic fibrosis transmembrane conductance regulator in pulmonary epithelial cell lines, ASB activity increased significantly (1). Also, we found marked reductions in arylsulfatases A and B, galactose-6-sulfatase, and steroid sulfatase in malignant mammary cell lines (MCF-7, T47D, and HCC 1937), compared with normal primary mammary epi...

mentioning

confidence: 63%

Section: Discussionmentioning

confidence: 89%

“…Malignant mammary tissue was frozen at Ϫ80°C after surgical excision. Normal mammary tissue was cultured as previously reported (2).…”

Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

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Distinct Effects of N-Acetylgalactosamine-4-sulfatase and Galactose-6-sulfatase Expression on Chondroitin Sulfates

Journal of Biological Chemistry

Kotlo

Shukla

et al. 2008

Self Cite

“…Measurements of ARSB Activity, Total Sulfated Glycosaminoglycans, and Chondroitin 4-Sulfate-ARSB was measured by a fluorometric assay, following a standard protocol using 20 l of homogenate and 80 l of assay buffer (0.05 M sodium acetate buffer, pH 5.6) with 100 l of substrate (5 mM 4-methylumbelliferyl sulfate in assay buffer) in a black microplate, as reported previously (33). Total sulfated glycosaminoglycans, including C4S, chondroitin 6-sulfate, keratan sulfate, dermatan sulfate, heparan sulfate, and heparin, were measured in NCM460 cells, HT-29 cells, mouse colon following exposure to carrageenan, and colon of ARSB-null mice by sulfated glycosaminoglycan (GAG) assay (Blyscan TM , Biocolor, Ltd., Newtownabbey, Northern Ireland) as described previously (32).…”

Section: Methodsmentioning

confidence: 99%

Increased Expression of Colonic Wnt9A through Sp1-mediated Transcriptional Effects involving Arylsulfatase B, Chondroitin 4-Sulfate, and Galectin-3

Journal of Biological Chemistry

Feferman

Tobacman

2014

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Background: Mechanism by which Wnt expression was increased by carrageenan exposure was unknown. Results: Sp1 activated Wnt9A transcription through changes in arylsulfatase B, chondroitin 4-sulfation, and galectin-3. Conclusion: Decline in arylsulfatase B leads to transcriptional effects mediated by Sp1 and galectin-3. Significance: Extracellular events can regulate transcription through changes in arylsulfatase B and chondroitin 4-sulfation.

Arylsulfatase B modulates neurite outgrowth via astrocyte chondroitin‐4‐sulfate: Dysregulation by ethanol

Zhang

Kusumo

et al. 2013

Glia

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In utero ethanol exposure causes Fetal Alcohol Spectrum Disorders, associated with reduced brain plasticity; the mechanisms of these effects are not well understood, particularly with respect to glial involvement. Astrocytes release factors that modulate neurite outgrowth. We explored the hypothesis that ethanol inhibits neurite outgrowth by increasing the release of inhibitory chondroitin sulfate proteoglycans (CSPGs) from astrocytes. Astrocyte treatment with ethanol inhibited the activity of arylsulfatase B (ARSB), the enzyme that removes sulfate groups from chondroitin-4-sulfate (C4S) and triggers the degradation of C4S, increased total sulfated glycosaminoglycans (GAGs), C4S, and neurocan core-protein content and inhibited neurite outgrowth in neurons co-cultured with ethanol-treated astrocytes in vitro, effects reversed by treatment with recombinant ARSB. Ethanol also inhibited ARSB activity and increased sulfate GAG and neurocan levels in the developing hippocampus after in vivo ethanol exposure. ARSB silencing increased the levels of sulfated GAGs, C4S, and neurocan in astrocytes and inhibited neurite outgrowth in co-cultured neurons, indicating that ARSB activity directly regulates C4S and affects neurocan expression. In summary, this study reports two major findings: ARSB modulates sulfated GAG and neurocan levels in astrocytes and astrocyte-mediated neurite outgrowth in co-cultured neurons; and ethanol inhibits the activity of ARSB, increases sulfated GAG, C4S, and neurocan levels, and thereby inhibits astrocyte-mediated neurite outgrowth. An unscheduled increase in CSPGs in the developing brain may lead to altered brain connectivity and to premature decrease in neuronal plasticity and therefore represents a novel mechanism by which ethanol can exert its neurodevelopmental effects.