title: Reduced sialylation status in UDP-N-acetylglucosamine-2-epimerase/N-acetylmannosamine kinase (GNE)-deficient mice.
abstract: Sialic acids are widely expressed as terminal carbohydrates on glycoconjugates of eukaryotic cells. They are involved in a variety of cellular functions, such as cell adhesion or signal recognition. The key enzyme of sialic acid biosynthesis is the bifunctional UDP-N-acetylglucosamine-2-epimerase/N-acetylmannosamine kinase (GNE), which catalyzes the first two steps of sialic acid biosynthesis in the cytosol. Previously, we have shown that inactivation of the GNE by gene targeting causes early embryonic lethality in mice, whereas heterozygous GNE-deficient mice are vital. In this study we compared the amount of membrane-bound sialic acids of wildtype mice with those of heterozygous GNE-deficient mice. For that we quantified membrane-bound sialic acid concentration in various organs of wildtype- and heterozygous GNE-deficient mice. We found an organ-specific reduction of membrane-bound sialic acids in heterozygous GNE-deficient mice. The overall reduction was 25%. Additionally, we analyzed transferrin and polysialylated neural cell adhesion molecule (NCAM) by one- or two-dimensional gel electrophoresis. Transferrin-expression was unchanged in heterozygous GNE-deficient mice; however the isoelectric point of transferrin was shifted towards basic pH, indicating a reduced sialylation. Furthermore, the expression of polysialic acids on NCAM was reduced in GNE-deficient mice.
authors.affiliation: Institut für Physiologische, Chemie Hollystr. 1, 06114, Halle (Saale), Germany.
authors.forename: Daniel
authors.initials: D
authors.lastname: Gagiannis
authors.forename: André
authors.initials: A
authors.lastname: Orthmann
authors.forename: Ilona
authors.initials: I
authors.lastname: Danssmann
authors.forename: Martina
authors.initials: M
authors.lastname: Schwarzkopf
authors.forename: Wenke
authors.initials: W
authors.lastname: Weidemann
authors.forename: Rüdiger
authors.initials: R
authors.lastname: Horstkorte
mesh_terms: D000818:Animals; D002462:Cell Membrane / Q000378:metabolism; D005260:Female; D006001:Glycoconjugates / Q000737:chemistry / Q000378:metabolism; D006579:Heterozygote; D051379:Mice; D008810:Mice, Inbred C57BL; D018345:Mice, Knockout; D009097:Multienzyme Complexes / Q000172:deficiency* / Q000235:genetics / Q000378:metabolism; D019006:Neural Cell Adhesion Molecules / Q000737:chemistry / Q000378:metabolism; D012794:Sialic Acids / Q000737:chemistry / Q000378:metabolism; D014018:Tissue Distribution; D014168:Transferrin / Q000737:chemistry / Q000378:metabolism
chemical_list: D006001:Glycoconjugates; D009097:Multienzyme Complexes; D019006:Neural Cell Adhesion Molecules; D012794:Sialic Acids; D014168:Transferrin; C482901:UDP-N-acetylglucosamine 2-epimerase - N-acetylmannosamine kinase