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.
Similar content being viewed by others
References
Varki, A.: Biological roles of oligosaccharides: all of the theories are correct. Glycobiology 3, 97 (1993)
Traving, C., Schauer, R.: Structure, function and metabolism of sialic acids. Cell. Mol. Life. Sci. 54, 1330 (1998)
Kelm, S., Schauer, R.: Sialic acids in molecular and cellular interactions. Int. Rev. Cytol. 175, 137 (1998)
Varki, A.: Sialic acids as ligands in recognition phenomena. Faseb. J. 11, 248 (1997)
Finne, J., Finne, U., Deagostini-Bazin, H., Goridis, C.: Occurrence of alpha 2–8 linked polysialosyl units in a neural cell adhesion molecule. Biochem. Biophys. Res. Commun. 112, 482 (1983)
Seki, T., Arai, Y.: Distribution and possible roles of the highly polysialylated neural cell adhesion molecule (NCAM-H) in the developing and adult central nervous system. Neurosci. Res. 17, 265 (1993)
Szele, F.G., Dowling, J.J., Gonzales, C., Theveniau, M., Rougon, G., Chesselet, M.F.: Pattern of expression of highly polysialylated neural cell adhesion molecule in the developing and adult rat striatum. Neuroscience 60, 133 (1994)
Daniel, L., Durbec, P., Gautherot, E., Rouvier, E., Rougon, G., Figarella-Branger, D.: A nude mice model of human rhabdomyosarcoma lung metastases for evaluating the role of polysialic acids in the metastatic process. Oncogene 20, 997 (2001)
Tanaka, F., Otake, Y., Nakagawa, T., Kawano, Y., Miyahara, R., Li, M., Yanagihara, K., Inui, K., Oyanagi, H., Yamada, T., Nakayama, J., Fujimoto, I., Ikenaka, K., Wada, H.: Prognostic significance of polysialic acid expression in resected non-small cell lung cancer. Cancer. Res. 15, 1666 (2001)
Stäsche, R., Hinderlich, S., Weise, C., Effertz, K., Lucka, L., Moormann, P., Reutter, W.: A bifunctional enzyme catalyzes the first two steps in N-acetylneuraminic acid biosynthesis of rat liver. Molecular cloning and functional expression of UDP-N-acetyl-glucosamine 2-epimerase/N-acetylmannosamine kinase. J. Biol. Chem. 272, 24319 (1997)
Hinderlich, S., Stäsche, R., Zeitler, R., Reutter, W.: A bifunctional enzyme catalyzes the first two steps in N-acetylneuraminic acid biosynthesis of rat liver. Purification and characterization of UDP-N-acetylglucosamine 2-epimerase/N-acetylmannosamine kinase. J. Biol. Chem. 272, 24313 (1997)
Krause, S., Hinderlich, S. Amsili, S., Horstkorte, R., Wiendl, H., Argov, Z., Mitrani-Rosenbaum, S., Lochmüller, H.: Localization of UDP-GlcNAc 2-epimerase/ManNAc (GNE) in the Golgi complex and the nucleus of mammalian cells. Exp. Cell. Res. 304, 365 (2005)
Horstkorte, R., Nöhring, S., Wiechens, N., Schwarzkopf, M., Danker, K., Reutter, W., Lucka, L.: Tissue expression and amino acid sequence of murine UDP-N-acetylglucosamine-2-epimerase/N-acetylmannosamine kinase. Eur. J. Biochem. 260, 923 (1999)
Horstkorte, R., Nöhring, S., Reutter, W., Effertz, K., Lucka, L., Danker, K.: Proteinkinase C phosphorylates and regulates UDP-N-acetylglucosamine-2-epimerase/N-acetyl-mannosamine kinase. FEBS Lett. 470, 315 (2000)
Seppala, R., Lehto, V.P., Gahl, W.A.: Mutations in the human UDP-N-acetyl-glucosamine 2-epimerase gene define the disease sialuria and the allosteric site of the enzyme. Am. J. Hum. Genet. 64, 1563 (1999)
Keppler, O.T., Hinderlich, S., Langner, J., Schwartz-Albiez, R., Reutter, W., Pawlita, M.: UDP-GlcNAc 2-epimerase: a regulator of cell surface sialylation. Science 284, 1372 (1999)
Eisenberg, I., Avidan, N., Potikha, T., Hochner, H., Chen, M., Olender, T., Barash, M., Shemesh, M., Sadeh, M., Grabov-Nardini, G., Shmilevich, I., Friedmann, A., Karpati, G., Bradley, W.G., Baumbach, L., Lancet, D., Asher, E.B., Beckmann, J.S., Argov, Z., Mitrani-Rosenbaum, S.: The UDP-N-acetylglucosamine 2-epimerase/N-acetylmannosamine kinase gene is mutated in recessive hereditary inclusion body myopathy. Nat. Genet. 29, 83–87 (2001)
Schwarzkopf, M., Knobeloch, K.P., Rohde, E., Hinderlich, S., Wiechens, N., Lucka, L., Horak, I., Reutter, W., Horstkorte, R.: Sialylation is essential for early development in mice. Proc. Natl. Acad. Sci. U S A 99, 5267 (2002)
Xin, Y., Lasker, J.M., Rosman, A.S., Lieber, C.S.: Isoelectric focusing/western blotting: a novel and practical method for quantitation of carbohydrate-deficient transferrin in alcoholics. Alcohol Clin. Exp. Res. 15, 814 (1991)
Eckhardt, M., Bukalo, O., Chazal, G., Wang, L., Goridis, C., Schachner, M., Gerardy-Schahn, R., Cremer, H., Dityatev, A.: Mice deficient in the polysialyltransferase ST8SiaIV/PST-1 allow discrimination of the roles of neural cell adhesion molecule protein and polysialic acid in neural development and synaptic plasticity. J. Neurosci. 20, 5234 (2000)
Angata, K., Long, J.M., Bukalo, O., Lee, W., Dityatev, A., Wynshaw-Boris, A., Schachner, M., Fukuda, M., Marth, J.D.: Sialyltransferase ST8Sia-II assembles a subset of polysialic acid that directs hippocampal axonal targeting and promotes fear behavior. J. Biol. Chem. 279, 32603 (2004)
Jourdian, G.W., Dean, L., Roseman, S.: The sialic acids. XI. A periodate-resorcinol method for the quantitative estimation of free sialic acids and their glycosides. J. Biol. Chem. 246, 430 (1971)
Löster, K., Kannicht, C.: 2D-electrophoresis: detection of glycosylation and influence an spot pattern. In: Kannicht, C. (ed.) Posttranslational Modifications of Proteins—Tools for Functional Proteomicsm. Humana, Totowa, New Jersey (2002)
Acknowledgements
We are grateful to Dr. C. Kannicht (Octapharma, Berlin) for help with the two-dimensional gel electrophoresis. This work received financial support by the Deutsche Forschungsgemeinschaft, the Hertie-Stiftung, the Sonnenfeld-Stiftung and the Fonds der Chemischen Industrie.
Author information
Authors and Affiliations
Corresponding author
Additional information
Daniel Gagiannis and André Orthmann have contributed equally to this work.
Rights and permissions
About this article
Cite this article
Gagiannis, D., Orthmann, A., Danßmann, I. et al. Reduced sialylation status in UDP-N-acetylglucosamine-2-epimerase/N-acetylmannosamine kinase (GNE)-deficient mice. Glycoconj J 24, 125–130 (2007). https://doi.org/10.1007/s10719-006-9019-7
Received:
Revised:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s10719-006-9019-7