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Efficient metabolic oligosaccharide engineering of glycoproteins by UDP-N-acetylglucosamine 2-epimerase/N-acetylmannosamine kinase (GNE) knock-down.
- Source :
-
Molecular bioSystems [Mol Biosyst] 2011 Jul; Vol. 7 (7), pp. 2245-51. Date of Electronic Publication: 2011 May 16. - Publication Year :
- 2011
-
Abstract
- Improving the accessibility and functions of therapeutic and diagnostic glycoproteins is one of the major goals of glycobiotechnology. Here we present that stable knock-down of UDP-N-acetylglucosamine 2-epimerase/N-acetylmannosamine kinase (GNE), the key enzyme in the sialic acid biosynthetic pathway, dramatically increases incorporation of N-acetylmannosamine analogues into glycoproteins of HEK293 cells. By means of these GNE-deficient cells highly sialylated glycoproteins can efficiently be decorated with reactive functional groups, which can be employed in bioorthogonal functionalization strategies for fluorescence labelling or biotinylation.
- Subjects :
- Antigens, CD metabolism
Carbohydrate Conformation
Carbohydrate Epimerases metabolism
Cell Adhesion Molecules metabolism
Glycoproteins chemistry
HEK293 Cells
Humans
N-Acetylneuraminic Acid metabolism
Oligosaccharides chemistry
Phosphotransferases (Alcohol Group Acceptor) deficiency
RNA, Small Interfering metabolism
Recombinant Proteins metabolism
Staining and Labeling
Gene Knockdown Techniques
Glycoproteins metabolism
Oligosaccharides metabolism
Phosphotransferases (Alcohol Group Acceptor) metabolism
Protein Engineering methods
Subjects
Details
- Language :
- English
- ISSN :
- 1742-2051
- Volume :
- 7
- Issue :
- 7
- Database :
- MEDLINE
- Journal :
- Molecular bioSystems
- Publication Type :
- Academic Journal
- Accession number :
- 21584309
- Full Text :
- https://doi.org/10.1039/c1mb05059a