Sulphate release from keratan sulphate and heparan sulphate by bovine N-acetylglucosamine-6-sulphate sulphatase

Ali Shilatifard; Richard D. Cummings

doi:10.1093/glycob/5.3.291

Sulphate release from keratan sulphate and heparan sulphate by bovine N-acetylglucosamine-6-sulphate sulphatase

Ali Shilatifard, Richard D. Cummings^*

^*Corresponding author for this work

Biochemistry and Molecular Genetics

Research output: Contribution to journal › Article › peer-review

4 Scopus citations

Abstract

The functions of sulphated monosaccharides within glycosaminoglycans (GAGs) and glycoproteins are being studied intensely, but progress is hindered by an inability to selectively desulphate glycoconjugates. We recently identified an N-acetylglucosamine-6-sulphate sulphatase (NG6SS) from bovine kidney that can remove sulphate from N-acetylglucosamine-6-sulphate (GlcNAc-6-SO₄) within oligosaccharides and glycoproteins. However, the potential 'endosulphatase' activity of the NG6SS toward GAGs is not known. To test for this possibility, [³H]glucosamine-, [³H]galactose- and ³⁵SO_4- labelled keratan sulphate (KS) were separately prepared by metabolic radiolabelling of bovine cornea. NG6SS quantitatively removed sulphate from KS without release of sugar fragments. The enzyme had a K_m of 4.7 mM toward free GlcNAc-6-SO₄, but its K_m for commercially available bovine corneal KS was found to be 9.1 μM. Analyses of both KS and heparan sulphate after treatment with NG6SS demonstrated significant loss of sulphate from GlcNAc-6-SO₄ in both GAGs. These findings may be relevant for future studies aimed at defining the function(s) of GlcNAc-6-SO₄ residues in GAGs and understanding the catabolism of GAGs, especially in regard to sulphatidoses, such as Sanfilippo D syndrome in humans, which involves a deficiency of NG6SS activity

Original language	English (US)
Pages (from-to)	291-297
Number of pages	7
Journal	Glycobiology
Volume	5
Issue number	3
DOIs	https://doi.org/10.1093/glycob/5.3.291
State	Published - May 1995

Funding

This work was supported by NIH grants CA37626 and Al 27135 to R.D.C. The authors thank Dr Rodger McEver and Patty Wilkins for helpful discussions during the course of this work. We also thank Ms Judy Gaar for help in preparing the manuscript.

Keywords

Catabolism
Endosulphatase
Glycosaminoglycans
Sulphation

ASJC Scopus subject areas

General Medicine

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10.1093/glycob/5.3.291

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@article{1f477fd6a8b3404c9bafc89c44dff201,

title = "Sulphate release from keratan sulphate and heparan sulphate by bovine N-acetylglucosamine-6-sulphate sulphatase",

abstract = "The functions of sulphated monosaccharides within glycosaminoglycans (GAGs) and glycoproteins are being studied intensely, but progress is hindered by an inability to selectively desulphate glycoconjugates. We recently identified an N-acetylglucosamine-6-sulphate sulphatase (NG6SS) from bovine kidney that can remove sulphate from N-acetylglucosamine-6-sulphate (GlcNAc-6-SO4) within oligosaccharides and glycoproteins. However, the potential 'endosulphatase' activity of the NG6SS toward GAGs is not known. To test for this possibility, [3H]glucosamine-, [3H]galactose- and 35SO4- labelled keratan sulphate (KS) were separately prepared by metabolic radiolabelling of bovine cornea. NG6SS quantitatively removed sulphate from KS without release of sugar fragments. The enzyme had a Km of 4.7 mM toward free GlcNAc-6-SO4, but its Km for commercially available bovine corneal KS was found to be 9.1 μM. Analyses of both KS and heparan sulphate after treatment with NG6SS demonstrated significant loss of sulphate from GlcNAc-6-SO4 in both GAGs. These findings may be relevant for future studies aimed at defining the function(s) of GlcNAc-6-SO4 residues in GAGs and understanding the catabolism of GAGs, especially in regard to sulphatidoses, such as Sanfilippo D syndrome in humans, which involves a deficiency of NG6SS activity",

keywords = "Catabolism, Endosulphatase, Glycosaminoglycans, Sulphation",

author = "Ali Shilatifard and Cummings, {Richard D.}",

note = "Funding Information: This work was supported by NIH grants CA37626 and Al 27135 to R.D.C. The authors thank Dr Rodger McEver and Patty Wilkins for helpful discussions during the course of this work. We also thank Ms Judy Gaar for help in preparing the manuscript.",

year = "1995",

month = may,

doi = "10.1093/glycob/5.3.291",

language = "English (US)",

volume = "5",

pages = "291--297",

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TY - JOUR

T1 - Sulphate release from keratan sulphate and heparan sulphate by bovine N-acetylglucosamine-6-sulphate sulphatase

AU - Shilatifard, Ali

AU - Cummings, Richard D.

N1 - Funding Information: This work was supported by NIH grants CA37626 and Al 27135 to R.D.C. The authors thank Dr Rodger McEver and Patty Wilkins for helpful discussions during the course of this work. We also thank Ms Judy Gaar for help in preparing the manuscript.

PY - 1995/5

Y1 - 1995/5

N2 - The functions of sulphated monosaccharides within glycosaminoglycans (GAGs) and glycoproteins are being studied intensely, but progress is hindered by an inability to selectively desulphate glycoconjugates. We recently identified an N-acetylglucosamine-6-sulphate sulphatase (NG6SS) from bovine kidney that can remove sulphate from N-acetylglucosamine-6-sulphate (GlcNAc-6-SO4) within oligosaccharides and glycoproteins. However, the potential 'endosulphatase' activity of the NG6SS toward GAGs is not known. To test for this possibility, [3H]glucosamine-, [3H]galactose- and 35SO4- labelled keratan sulphate (KS) were separately prepared by metabolic radiolabelling of bovine cornea. NG6SS quantitatively removed sulphate from KS without release of sugar fragments. The enzyme had a Km of 4.7 mM toward free GlcNAc-6-SO4, but its Km for commercially available bovine corneal KS was found to be 9.1 μM. Analyses of both KS and heparan sulphate after treatment with NG6SS demonstrated significant loss of sulphate from GlcNAc-6-SO4 in both GAGs. These findings may be relevant for future studies aimed at defining the function(s) of GlcNAc-6-SO4 residues in GAGs and understanding the catabolism of GAGs, especially in regard to sulphatidoses, such as Sanfilippo D syndrome in humans, which involves a deficiency of NG6SS activity

AB - The functions of sulphated monosaccharides within glycosaminoglycans (GAGs) and glycoproteins are being studied intensely, but progress is hindered by an inability to selectively desulphate glycoconjugates. We recently identified an N-acetylglucosamine-6-sulphate sulphatase (NG6SS) from bovine kidney that can remove sulphate from N-acetylglucosamine-6-sulphate (GlcNAc-6-SO4) within oligosaccharides and glycoproteins. However, the potential 'endosulphatase' activity of the NG6SS toward GAGs is not known. To test for this possibility, [3H]glucosamine-, [3H]galactose- and 35SO4- labelled keratan sulphate (KS) were separately prepared by metabolic radiolabelling of bovine cornea. NG6SS quantitatively removed sulphate from KS without release of sugar fragments. The enzyme had a Km of 4.7 mM toward free GlcNAc-6-SO4, but its Km for commercially available bovine corneal KS was found to be 9.1 μM. Analyses of both KS and heparan sulphate after treatment with NG6SS demonstrated significant loss of sulphate from GlcNAc-6-SO4 in both GAGs. These findings may be relevant for future studies aimed at defining the function(s) of GlcNAc-6-SO4 residues in GAGs and understanding the catabolism of GAGs, especially in regard to sulphatidoses, such as Sanfilippo D syndrome in humans, which involves a deficiency of NG6SS activity

KW - Catabolism

KW - Endosulphatase

KW - Glycosaminoglycans

KW - Sulphation

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AN - SCOPUS:0029034514

SN - 0959-6658

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SP - 291

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JO - Glycobiology

JF - Glycobiology

IS - 3

ER -

Sulphate release from keratan sulphate and heparan sulphate by bovine N-acetylglucosamine-6-sulphate sulphatase

Abstract

Funding

Keywords

ASJC Scopus subject areas

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