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

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.

• Authors: Kaya Bork, Werner Reutter, Wenke Weidemann, Rüdiger Horstkorte
• Journal: FEBS letters (FEBS Lett) Vol. 581 Issue 22 Pg. 4195-8 (Sep 04 2007) ISSN: 0014-5793 [Print] England
• PMID: 17706199 (Publication Type: Journal Article, Research Support, Non-U.S. Gov't)

Chemical References

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

Topics

• 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, metabolism)
• Rats
• Recombinant Proteins
• Sialic Acid Storage Disease (enzymology)