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De-N-glycosylation or G82S mutation of RAGE sensitizes its interaction with advanced glycation endproducts

Mari Osawa1, Yasuhiko Yamamoto, Seiichi Munesue

  • 1Department of Biochemistry and Molecular Vascular Biology, Kanazawa University Graduate School of Medical Science, 13-1 Takara-machi, Kanazawa 920-8640, Japan.

Biochimica Et Biophysica Acta
|August 24, 2007
PubMed

Insights

De-N-glycosylation or G82S mutation of the receptor for advanced glycation endproducts (RAGE) enhances its AGE binding affinity. This modification may sensitize cells to AGE, potentially impacting diabetic vascular complications.

Area of Science:

  • Biochemistry
  • Molecular Biology
  • Cell Biology

Background:

  • Advanced glycation endproducts (AGE) and their receptor (RAGE) interactions are linked to diabetic vascular complications.
  • RAGE possesses N-glycosylation sites near its AGE-binding domain, with a G82S mutation identified in humans.

Purpose of the Study:

  • To investigate the impact of RAGE de-N-glycosylation and G82S mutation on AGE binding.
  • To assess cellular responses to AGE in cells expressing modified RAGE.

Main Methods:

  • Production and purification of recombinant wild-type, de-N-glycosylated, and G82S RAGE proteins in COS-7 cells.
  • Ligand-binding assays to determine dissociation constants (Kd).
  • Analysis of VEGF mRNA expression in ECV304 cells under varying RAGE expression and glucose conditions.

Main Results:

  • De-N-glycosylation and G82S mutation significantly decreased the Kd for glycolaldehyde-derived AGE by three orders of magnitude.
  • AGE-induced VEGF mRNA upregulation was augmented in cells expressing de-N-glycosylated or G82S RAGE.
  • Low glucose conditions induced RAGE deglycosylation and enhanced AGE-induced VEGF mRNA expression.

Conclusions:

  • RAGE de-N-glycosylation or G82S mutation increases affinity for AGE ligands.
  • These modifications may sensitize cells to AGE, potentially contributing to diabetic vascular complications.

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