title: 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: Institut für Biochemie und Molekularbiologie, Charité-Universitätsmedizin Berlin, Campus Benjamin Franklin, Arnimallee 22, 14195 Berlin-Dahlem, Germany.
authors: Kaya
authors: K
authors: Bork
authors: Werner
authors: W
authors: Reutter
authors: Wenke
authors: W
authors: Weidemann
authors: Rüdiger
authors: R
authors: Horstkorte
mesh_terms: D000818:Animals; D001973:Bromodeoxyuridine / Q000378:metabolism; D016466:CHO Cells; D002238:Carbohydrate Epimerases / Q000235:genetics; D049109:Cell Proliferation; D006224:Cricetinae; D003412:Cricetulus; D017077:Culture Media, Conditioned; D004247:DNA / Q000096:biosynthesis; D004921:Erythropoietin / Q000378:metabolism*; D006801:Humans; D007525:Isoelectric Focusing; D008954:Models, Biological; D009154:Mutation / Q000235:genetics*; D019158:N-Acetylneuraminic Acid / Q000032:analysis / Q000378:metabolism*; D051381:Rats; D011994:Recombinant Proteins; D029461:Sialic Acid Storage Disease / Q000201:enzymology*
chemical_list: D017077:Culture Media, Conditioned; D011994:Recombinant Proteins; D004921:Erythropoietin; D004247:DNA; D002238:Carbohydrate Epimerases; C100033:UDP acetylglucosamine-2-epimerase; D001973:Bromodeoxyuridine; D019158:N-Acetylneuraminic Acid