5-thiomannosides block the biosynthesis of dolichol-linked oligosaccharides and mimic class I congenital disorders of glycosylation

Wesley F. Zandberg, Ningguo Gao, Jayakanthan Kumarasamy, Mark A. Lehrman, Nabil G. Seidah, B. Mario Pinto

Research output: Contribution to journalArticle

2 Citations (Scopus)

Abstract

In a cell-based assay for novel inhibitors, we have discovered that two glycosides of 5-thiomannose, each containing an interglycosidic nitrogen atom, prevented the correct zymogen processing of the prohormone proopiomelanocortinin (POMC) and the transcription factor sterol-regulatory element-binding protein-2 (SREBP-2) in mouse pituitary cells and Chinese hamster ovary (CHO) cells, respectively. In the case of SREBP-2, these effects were correlated with the altered N-linked glycosylation of subtilisin/kexin-like isozyme-1 (SKI-1), the protease responsible for SREBP-2 processing under sterol-limiting conditions. Further examination of the effects of these compounds in CHO cells showed that they cause extensive protein hypoglycosylation in a manner similar to type I congenital disorders of glycosylation (CDGs) since the remaining N-glycans in treated cells were complete (normal) structures. The under-glycosylation of glycoproteins in 5-thiomannoside-treated cells is now shown to be caused by the compromised biosynthesis of the dolichol-linked oligosaccharide (DLO) N-glycosylation donor, although the nucleotide sugars required for the synthesis of DLOs were neither reduced under these conditions, nor were their effects reversed upon the addition of exogenous mannose. Analysis of DLO intermediates by fluorophore-assisted carbohydrate electrophoresis demonstrated that 5-thiomannose-containing glycosides block DLO biosynthesis most likely at a stage prior to the GlcNAc 2Man 3 intermediate, on the cytosolic face of the endoplasmic reticulum.

Original languageEnglish (US)
Pages (from-to)392-401
Number of pages10
JournalChemBioChem
Volume13
Issue number3
DOIs
StatePublished - Feb 13 2012

Fingerprint

Congenital Disorders of Glycosylation
Dolichol
Glycosylation
Sterol Regulatory Element Binding Protein 2
Biosynthesis
Oligosaccharides
Cells
Glycosides
Cricetulus
Subtilisin
Enzyme Precursors
Fluorophores
Ovary
Sterols
Mannose
Processing
Electrophoresis
Sugars
Isoenzymes
Polysaccharides

Keywords

  • 5-thiomannosides
  • Glycoproteins
  • Mannosyltransferases
  • Nucleotide sugars
  • Oligosaccharides

ASJC Scopus subject areas

  • Biochemistry
  • Organic Chemistry
  • Molecular Medicine
  • Molecular Biology

Cite this

5-thiomannosides block the biosynthesis of dolichol-linked oligosaccharides and mimic class I congenital disorders of glycosylation. / Zandberg, Wesley F.; Gao, Ningguo; Kumarasamy, Jayakanthan; Lehrman, Mark A.; Seidah, Nabil G.; Pinto, B. Mario.

In: ChemBioChem, Vol. 13, No. 3, 13.02.2012, p. 392-401.

Research output: Contribution to journalArticle

Zandberg, Wesley F. ; Gao, Ningguo ; Kumarasamy, Jayakanthan ; Lehrman, Mark A. ; Seidah, Nabil G. ; Pinto, B. Mario. / 5-thiomannosides block the biosynthesis of dolichol-linked oligosaccharides and mimic class I congenital disorders of glycosylation. In: ChemBioChem. 2012 ; Vol. 13, No. 3. pp. 392-401.
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AU - Kumarasamy, Jayakanthan

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AU - Seidah, Nabil G.

AU - Pinto, B. Mario

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AB - In a cell-based assay for novel inhibitors, we have discovered that two glycosides of 5-thiomannose, each containing an interglycosidic nitrogen atom, prevented the correct zymogen processing of the prohormone proopiomelanocortinin (POMC) and the transcription factor sterol-regulatory element-binding protein-2 (SREBP-2) in mouse pituitary cells and Chinese hamster ovary (CHO) cells, respectively. In the case of SREBP-2, these effects were correlated with the altered N-linked glycosylation of subtilisin/kexin-like isozyme-1 (SKI-1), the protease responsible for SREBP-2 processing under sterol-limiting conditions. Further examination of the effects of these compounds in CHO cells showed that they cause extensive protein hypoglycosylation in a manner similar to type I congenital disorders of glycosylation (CDGs) since the remaining N-glycans in treated cells were complete (normal) structures. The under-glycosylation of glycoproteins in 5-thiomannoside-treated cells is now shown to be caused by the compromised biosynthesis of the dolichol-linked oligosaccharide (DLO) N-glycosylation donor, although the nucleotide sugars required for the synthesis of DLOs were neither reduced under these conditions, nor were their effects reversed upon the addition of exogenous mannose. Analysis of DLO intermediates by fluorophore-assisted carbohydrate electrophoresis demonstrated that 5-thiomannose-containing glycosides block DLO biosynthesis most likely at a stage prior to the GlcNAc 2Man 3 intermediate, on the cytosolic face of the endoplasmic reticulum.

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