Enhanced sialylation of EPO by overexpression of UDP-GlcNAc 2-epimerase/ManAc kinase containing a sialuria mutation in CHO cells

FEBS Lett. 2007 Sep 4;581(22):4195-8. doi: 10.1016/j.febslet.2007.07.060. Epub 2007 Aug 3.

Abstract

Sialylation (e.g. expression of sialic acid) plays a crucial role for function and stability of most glycoproteins. The key enzyme for the biosynthesis of sialic acid is the UDP-N-acetylglucosamine 2-epimerase/N-acetylmannosamine-kinase (GNE). Mutations in the binding site of the feedback inhibitor CMP-sialic acid of the GNE leads to sialuria, a disease in which patients produce sialic acid in gram scale. Here, we report on the use in biotechnology of sialuria-mutated GNE. Expression of the sialuria-mutated GNE in CHO-cells leads to increased sialylation of recombinant expressed erythropoietin (EPO). Our data show that sialuria-mutated-GNE over-expressing cells are the perfect platform to express highly sialylated therapeutic proteins, such as EPO.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Bromodeoxyuridine / metabolism
  • CHO Cells
  • Carbohydrate Epimerases / genetics
  • Cell Proliferation
  • Cricetinae
  • Cricetulus
  • Culture Media, Conditioned
  • DNA / biosynthesis
  • Erythropoietin / metabolism*
  • Humans
  • Isoelectric Focusing
  • Models, Biological
  • Mutation / genetics*
  • N-Acetylneuraminic Acid / analysis
  • N-Acetylneuraminic Acid / metabolism*
  • Rats
  • Recombinant Proteins
  • Sialic Acid Storage Disease / enzymology*

Substances

  • Culture Media, Conditioned
  • Recombinant Proteins
  • Erythropoietin
  • DNA
  • Carbohydrate Epimerases
  • UDP acetylglucosamine-2-epimerase
  • Bromodeoxyuridine
  • N-Acetylneuraminic Acid