Selective depolymerisation of dermatan sulfate: Production of radiolabelled substrates for α-l-iduronidase, sulfoiduronate sulfatase, and β-d-glucuronidase

John J. Hopwood, Vivienne J. Muller

Research output: Contribution to journalArticle

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Abstract

Radiolabelled disaccharide substrates for α-l-iduronidase, β-d-glucuronidase, and sulfoiduronate sulfatase have been prepared from dermatan sulfate by application in sequence of N-deacetylation, deaminative cleavage, and reduction with NaBT4. The yield of disaccharides was ∼87% of the total oligosaccharide fraction. Five disaccharides were isolated and tentatively identified. The major disaccharide, O-(α-l-idopyranosyluronic acid)-(1→3)-2,5-anhydro-d-[1-3H]talitol 4-sulfate (IdoA-anT4S), represented ∼75% of the total disaccharide fraction. The other disaccharides were O-(α-l-idopyranosyluronic acid 2-sulfate)-(1→3)-2,5-anhydro-d-[1-3H]talitol 4-sulfate (IdoA2S-anT4S), O-(β-d-glucopyranosyluronic acid)-(1→3)-2,5-anhydro-d-[1-3H]talitol 4-sulfate (GlcA-anT4S), O-(β-d-glucopyranosyluronic acid)-(1→3)-2,5-anhydro-d-[1-3H]talitol 6-sulfate (GlcA-anT6S), and O-(α-l-idopyranosyluronic acid)-(1→3)-2,5-anhydro-d-[1-3H]talitol (IdoA-anT), which represented ∼4.5, 11.2, 1.0, and 1.8%, respectively, of the total disaccharide fraction. When incubated with cultured skin-fibroblasts from normal controls, IdoA-anT4S was shown to be a sensitive substrate for α-l-iduronidase to produce 2,5-anhydro-d-talitol 4-sulfate (anT4S). Activity toward IdoA-anT4S was not observed with fibroblast homogenates from α-l-iduronidase-deficient patients (Mucopolysaccharidosis Type I). Similarly, normal-fibroblast homogenates degraded GlcA-anT6S to anT6S, and GlcA-anT4S to anT4S, at a rate 6 to 8 times greater than found for fibroblasts from β-d-glucuronidase-deficient patients (Mucopolysaccharidosis Type VII). IdoA-anT4S was hydrolysed by α-l-iduronidase at a rate 365 times greater than that for IdoA-anT. Sulfation of the anhydro-d-[1-3H]talitol residues is an important structural determinant in the mechanism of action of α-l-iduronidase on disaccharide substrates. IdoA2S-anT4S was degraded to IdoA-anT4S and then to anT4S by normal-fibroblast homogenates, whereas fibroblasts from α-l-iduronidase-deficient and sulfoiduronate sulfatase-deficient (Mucopolysaccharidosis Type II) patients produced considerably decreased levels of anT4s and IdoA-anT4S (and anT4S), respectively.

LanguageEnglish
Pages227-239
Number of pages13
JournalCarbohydrate Research
Volume122
Issue number2
DOIs
Publication statusPublished - 28 Oct 1983

ASJC Scopus subject areas

  • Analytical Chemistry
  • Biochemistry
  • Organic Chemistry

Cite this

@article{51bd8cea47a44743aa66aea79ef54034,
title = "Selective depolymerisation of dermatan sulfate: Production of radiolabelled substrates for α-l-iduronidase, sulfoiduronate sulfatase, and β-d-glucuronidase",
abstract = "Radiolabelled disaccharide substrates for α-l-iduronidase, β-d-glucuronidase, and sulfoiduronate sulfatase have been prepared from dermatan sulfate by application in sequence of N-deacetylation, deaminative cleavage, and reduction with NaBT4. The yield of disaccharides was ∼87{\%} of the total oligosaccharide fraction. Five disaccharides were isolated and tentatively identified. The major disaccharide, O-(α-l-idopyranosyluronic acid)-(1→3)-2,5-anhydro-d-[1-3H]talitol 4-sulfate (IdoA-anT4S), represented ∼75{\%} of the total disaccharide fraction. The other disaccharides were O-(α-l-idopyranosyluronic acid 2-sulfate)-(1→3)-2,5-anhydro-d-[1-3H]talitol 4-sulfate (IdoA2S-anT4S), O-(β-d-glucopyranosyluronic acid)-(1→3)-2,5-anhydro-d-[1-3H]talitol 4-sulfate (GlcA-anT4S), O-(β-d-glucopyranosyluronic acid)-(1→3)-2,5-anhydro-d-[1-3H]talitol 6-sulfate (GlcA-anT6S), and O-(α-l-idopyranosyluronic acid)-(1→3)-2,5-anhydro-d-[1-3H]talitol (IdoA-anT), which represented ∼4.5, 11.2, 1.0, and 1.8{\%}, respectively, of the total disaccharide fraction. When incubated with cultured skin-fibroblasts from normal controls, IdoA-anT4S was shown to be a sensitive substrate for α-l-iduronidase to produce 2,5-anhydro-d-talitol 4-sulfate (anT4S). Activity toward IdoA-anT4S was not observed with fibroblast homogenates from α-l-iduronidase-deficient patients (Mucopolysaccharidosis Type I). Similarly, normal-fibroblast homogenates degraded GlcA-anT6S to anT6S, and GlcA-anT4S to anT4S, at a rate 6 to 8 times greater than found for fibroblasts from β-d-glucuronidase-deficient patients (Mucopolysaccharidosis Type VII). IdoA-anT4S was hydrolysed by α-l-iduronidase at a rate 365 times greater than that for IdoA-anT. Sulfation of the anhydro-d-[1-3H]talitol residues is an important structural determinant in the mechanism of action of α-l-iduronidase on disaccharide substrates. IdoA2S-anT4S was degraded to IdoA-anT4S and then to anT4S by normal-fibroblast homogenates, whereas fibroblasts from α-l-iduronidase-deficient and sulfoiduronate sulfatase-deficient (Mucopolysaccharidosis Type II) patients produced considerably decreased levels of anT4s and IdoA-anT4S (and anT4S), respectively.",
author = "Hopwood, {John J.} and Muller, {Vivienne J.}",
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month = "10",
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language = "English",
volume = "122",
pages = "227--239",
journal = "Carbohydrate Research",
issn = "0008-6215",
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}

Selective depolymerisation of dermatan sulfate : Production of radiolabelled substrates for α-l-iduronidase, sulfoiduronate sulfatase, and β-d-glucuronidase. / Hopwood, John J.; Muller, Vivienne J.

In: Carbohydrate Research, Vol. 122, No. 2, 28.10.1983, p. 227-239.

Research output: Contribution to journalArticle

TY - JOUR

T1 - Selective depolymerisation of dermatan sulfate

T2 - Carbohydrate Research

AU - Hopwood, John J.

AU - Muller, Vivienne J.

PY - 1983/10/28

Y1 - 1983/10/28

N2 - Radiolabelled disaccharide substrates for α-l-iduronidase, β-d-glucuronidase, and sulfoiduronate sulfatase have been prepared from dermatan sulfate by application in sequence of N-deacetylation, deaminative cleavage, and reduction with NaBT4. The yield of disaccharides was ∼87% of the total oligosaccharide fraction. Five disaccharides were isolated and tentatively identified. The major disaccharide, O-(α-l-idopyranosyluronic acid)-(1→3)-2,5-anhydro-d-[1-3H]talitol 4-sulfate (IdoA-anT4S), represented ∼75% of the total disaccharide fraction. The other disaccharides were O-(α-l-idopyranosyluronic acid 2-sulfate)-(1→3)-2,5-anhydro-d-[1-3H]talitol 4-sulfate (IdoA2S-anT4S), O-(β-d-glucopyranosyluronic acid)-(1→3)-2,5-anhydro-d-[1-3H]talitol 4-sulfate (GlcA-anT4S), O-(β-d-glucopyranosyluronic acid)-(1→3)-2,5-anhydro-d-[1-3H]talitol 6-sulfate (GlcA-anT6S), and O-(α-l-idopyranosyluronic acid)-(1→3)-2,5-anhydro-d-[1-3H]talitol (IdoA-anT), which represented ∼4.5, 11.2, 1.0, and 1.8%, respectively, of the total disaccharide fraction. When incubated with cultured skin-fibroblasts from normal controls, IdoA-anT4S was shown to be a sensitive substrate for α-l-iduronidase to produce 2,5-anhydro-d-talitol 4-sulfate (anT4S). Activity toward IdoA-anT4S was not observed with fibroblast homogenates from α-l-iduronidase-deficient patients (Mucopolysaccharidosis Type I). Similarly, normal-fibroblast homogenates degraded GlcA-anT6S to anT6S, and GlcA-anT4S to anT4S, at a rate 6 to 8 times greater than found for fibroblasts from β-d-glucuronidase-deficient patients (Mucopolysaccharidosis Type VII). IdoA-anT4S was hydrolysed by α-l-iduronidase at a rate 365 times greater than that for IdoA-anT. Sulfation of the anhydro-d-[1-3H]talitol residues is an important structural determinant in the mechanism of action of α-l-iduronidase on disaccharide substrates. IdoA2S-anT4S was degraded to IdoA-anT4S and then to anT4S by normal-fibroblast homogenates, whereas fibroblasts from α-l-iduronidase-deficient and sulfoiduronate sulfatase-deficient (Mucopolysaccharidosis Type II) patients produced considerably decreased levels of anT4s and IdoA-anT4S (and anT4S), respectively.

AB - Radiolabelled disaccharide substrates for α-l-iduronidase, β-d-glucuronidase, and sulfoiduronate sulfatase have been prepared from dermatan sulfate by application in sequence of N-deacetylation, deaminative cleavage, and reduction with NaBT4. The yield of disaccharides was ∼87% of the total oligosaccharide fraction. Five disaccharides were isolated and tentatively identified. The major disaccharide, O-(α-l-idopyranosyluronic acid)-(1→3)-2,5-anhydro-d-[1-3H]talitol 4-sulfate (IdoA-anT4S), represented ∼75% of the total disaccharide fraction. The other disaccharides were O-(α-l-idopyranosyluronic acid 2-sulfate)-(1→3)-2,5-anhydro-d-[1-3H]talitol 4-sulfate (IdoA2S-anT4S), O-(β-d-glucopyranosyluronic acid)-(1→3)-2,5-anhydro-d-[1-3H]talitol 4-sulfate (GlcA-anT4S), O-(β-d-glucopyranosyluronic acid)-(1→3)-2,5-anhydro-d-[1-3H]talitol 6-sulfate (GlcA-anT6S), and O-(α-l-idopyranosyluronic acid)-(1→3)-2,5-anhydro-d-[1-3H]talitol (IdoA-anT), which represented ∼4.5, 11.2, 1.0, and 1.8%, respectively, of the total disaccharide fraction. When incubated with cultured skin-fibroblasts from normal controls, IdoA-anT4S was shown to be a sensitive substrate for α-l-iduronidase to produce 2,5-anhydro-d-talitol 4-sulfate (anT4S). Activity toward IdoA-anT4S was not observed with fibroblast homogenates from α-l-iduronidase-deficient patients (Mucopolysaccharidosis Type I). Similarly, normal-fibroblast homogenates degraded GlcA-anT6S to anT6S, and GlcA-anT4S to anT4S, at a rate 6 to 8 times greater than found for fibroblasts from β-d-glucuronidase-deficient patients (Mucopolysaccharidosis Type VII). IdoA-anT4S was hydrolysed by α-l-iduronidase at a rate 365 times greater than that for IdoA-anT. Sulfation of the anhydro-d-[1-3H]talitol residues is an important structural determinant in the mechanism of action of α-l-iduronidase on disaccharide substrates. IdoA2S-anT4S was degraded to IdoA-anT4S and then to anT4S by normal-fibroblast homogenates, whereas fibroblasts from α-l-iduronidase-deficient and sulfoiduronate sulfatase-deficient (Mucopolysaccharidosis Type II) patients produced considerably decreased levels of anT4s and IdoA-anT4S (and anT4S), respectively.

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