HOME HELP FEEDBACK SUBSCRIPTIONS ARCHIVE SEARCH TABLE OF CONTENTS		QUICK SEARCH:   [advanced] Author: Keyword(s): Year:  Vol:  Page:

This Article Full Text Full Text (PDF) Alert me when this article is cited Alert me if a correction is posted Citation Map Services Email this article to a friend Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Citing Articles Citing Articles via HighWire Citing Articles via Google Scholar Google Scholar Articles by Stäsche, R. Articles by Reutter, W. Search for Related Content PubMed PubMed Citation Articles by Stäsche, R. Articles by Reutter, W. Social Bookmarking                      What's this?

Volume 272, Number 39, Issue of September 26, 1997 pp. 24319-24324
©1997 by The American Society for Biochemistry and Molecular Biology, Inc.

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

(Received for publication, June 3, 1997)

Roger Stäsche , Stephan Hinderlich , Christoph Weise ^§ , Karin Effertz , Lothar Lucka , Petra Moormann and Werner Reutter

From the  Institut für Molekularbiologie und Biochemie, Freie Universität Berlin, Arnimallee 22, D-14195 Berlin-Dahlem, Germany and the ^§ Institut für Biochemie, Freie Universität Berlin, Thielallee 63, D-14195 Berlin-Dahlem, Germany

N-Acetylneuraminic acid (Neu5Ac) is the precursor of sialic acids, a group of important molecules in biological recognition systems. Biosynthesis of Neu5Ac is initiated and regulated by its key enzyme, UDP-N-acetylglucosamine 2-epimerase (UDP-GlcNAc 2-epimerase, EC 5.1.3.14)/N-acetylmannosamine kinase (ManNAc kinase, EC 2.7.1.60) in rat liver (Hinderlich, S., Stäsche, R., Zeitler, R., and Reutter, W. (1997) J. Biol. Chem. 272, 24313-24318). In the present paper we report the isolation and characterization of a cDNA clone encoding this bifunctional enzyme. An open reading frame of 2166 base pairs encodes 722 amino acids with a predicted molecular mass of 79 kDa. The deduced amino acid sequence contains exact matches of the sequences of five peptides derived from tryptic cleavage of the enzyme. The recombinant bifunctional enzyme was expressed in COS7 cells, where it displayed both epimerase and kinase activity.

Distribution of UDP-GlcNAc 2-epimerase/ManNAc kinase in the cytosol of several rat tissues was investigated by determining both specific enzyme activities. Secreting organs (liver, salivary glands, and intestinal mucosa) showed high specific activities of UDP-GlcNAc 2-epimerase/ManNAc kinase, whereas significant levels of these activities were absent from other organs (lung, kidney, spleen, brain, heart, skeletal muscle, and testis). Northern blot analysis revealed no UDP-GlcNAc 2-epimerase/ManNAc kinase mRNA in the non-secreting tissues.

CiteULike    Complore    Connotea    Del.icio.us    Digg    Reddit    Technorati    What's this?

This article has been cited by other articles:

				L-S Ro, G-J Lee-Chen, Y-R Wu, M Lee, P-Y Hsu, and C-M Chen Phenotypic variability in a Chinese family with rimmed vacuolar distal myopathy J. Neurol. Neurosurg. Psychiatry, May 1, 2005; 76(5): 752 - 755. [Abstract] [Full Text] [PDF]

				Y. Tajima, E. Uyama, S. Go, C. Sato, N. Tao, M. Kotani, H. Hino, A. Suzuki, Y. Sanai, K. Kitajima, et al. Distal Myopathy with Rimmed Vacuoles: Impaired O-Glycan Formation in Muscular Glycoproteins Am. J. Pathol., April 1, 2005; 166(4): 1121 - 1130. [Abstract] [Full Text] [PDF]

				A. Blume, A. J. Benie, F. Stolz, R. R. Schmidt, W. Reutter, S. Hinderlich, and T. Peters Characterization of Ligand Binding to the Bifunctional Key Enzyme in the Sialic Acid Biosynthesis by NMR: I. INVESTIGATION OF THE UDP-GlcNAc 2-EPIMERASE FUNCTIONALITY J. Biol. Chem., December 31, 2004; 279(53): 55715 - 55721. [Abstract] [Full Text] [PDF]

				A. J. Benie, A. Blume, R. R. Schmidt, W. Reutter, S. Hinderlich, and T. Peters Characterization of Ligand Binding to the Bifunctional Key Enzyme in the Sialic Acid Biosynthesis by NMR: II. INVESTIGATION OF THE ManNAc KINASE FUNCTIONALITY J. Biol. Chem., December 31, 2004; 279(53): 55722 - 55727. [Abstract] [Full Text] [PDF]

				A. K. Munster-Kuhnel, J. Tiralongo, S. Krapp, B. Weinhold, V. Ritz-Sedlacek, U. Jacob, and R. Gerardy-Schahn Structure and function of vertebrate CMP-sialic acid synthetases Glycobiology, October 1, 2004; 14(10): 43R - 51R. [Abstract] [Full Text] [PDF]

				Y. Hong and P. Stanley Lec3 Chinese Hamster Ovary Mutants Lack UDP-N-acetylglucosamine 2-Epimerase Activity Because of Mutations in the Epimerase Domain of the Gne Gene J. Biol. Chem., December 26, 2003; 278(52): 53045 - 53054. [Abstract] [Full Text] [PDF]

				K. Shinzawa and Y. Tsujimoto PLA2 activity is required for nuclear shrinkage in caspase-independent cell death J. Cell Biol., December 22, 2003; 163(6): 1219 - 1230. [Abstract] [Full Text] [PDF]

				J. J. Aumiller, J. R. Hollister, and D. L. Jarvis A transgenic insect cell line engineered to produce CMP-sialic acid and sialylated glycoproteins Glycobiology, June 1, 2003; 13(6): 497 - 507. [Abstract] [Full Text] [PDF]

				V. Askanas and W. King Engel Unfolding Story of Inclusion-Body Myositis and Myopathies: Role of Misfolded Proteins, Amyloid-{beta}, Cholesterol, and Aging J Child Neurol, March 1, 2003; 18(3): 185 - 190. [Abstract] [PDF]

				S. J. Luchansky, K. J. Yarema, S. Takahashi, and C. R. Bertozzi GlcNAc 2-Epimerase Can Serve a Catabolic Role in Sialic Acid Metabolism J. Biol. Chem., February 28, 2003; 278(10): 8035 - 8042. [Abstract] [Full Text] [PDF]

				H. Tomimitsu, K. Ishikawa, J. Shimizu, N. Ohkoshi, I. Kanazawa, and H. Mizusawa Distal myopathy with rimmed vacuoles: Novel mutations in the GNE gene Neurology, August 13, 2002; 59(3): 451 - 454. [Abstract] [Full Text] [PDF]

				M. Schwarzkopf, K.-P. Knobeloch, E. Rohde, S. Hinderlich, N. Wiechens, L. Lucka, I. Horak, W. Reutter, and R. Horstkorte Sialylation is essential for early development in mice PNAS, April 16, 2002; 99(8): 5267 - 5270. [Abstract] [Full Text] [PDF]

				H. Chen, A. Blume, M. Zimmermann-Kordmann, W. Reutter, and S. Hinderlich Purification and characterization of N-acetylneuraminic acid-9-phosphate synthase from rat liver Glycobiology, February 1, 2002; 12(2): 65 - 71. [Abstract] [Full Text] [PDF]

				M. Ringenberg, C. Lichtensteiger, and E. Vimr Redirection of sialic acid metabolism in genetically engineered Escherichia coli Glycobiology, July 1, 2001; 11(7): 533 - 539. [Abstract] [Full Text] [PDF]

				K. Effertz, S. Hinderlich, and W. Reutter Selective Loss of either the Epimerase or Kinase Activity of UDP-N-acetylglucosamine 2-Epimerase/N-Acetylmannosamine Kinase due to Site-directed Mutagenesis Based on Sequence Alignments J. Biol. Chem., October 1, 1999; 274(40): 28771 - 28778. [Abstract] [Full Text] [PDF]

				T. Angata, D. Nakata, T. Matsuda, K. Kitajima, and F. A. Troy II Biosynthesis of KDN (2-Keto-3-deoxy-D-glycero-D-galacto-nononic acid). IDENTIFICATION AND CHARACTERIZATION OF A KDN-9-PHOSPHATE SYNTHETASE ACTIVITY FROM TROUT TESTIS J. Biol. Chem., August 13, 1999; 274(33): 22949 - 22956. [Abstract] [Full Text] [PDF]

				O. T. Keppler, S. Hinderlich, J. Langner, R. Schwartz-Albiez, W. Reutter, and M. Pawlita UDP-GlcNAc 2-Epimerase: A Regulator of Cell Surface Sialylation Science, May 21, 1999; 284(5418): 1372 - 1376. [Abstract] [Full Text]

				J. Plumbridge and E. Vimr Convergent Pathways for Utilization of the Amino Sugars N-Acetylglucosamine, N-Acetylmannosamine, and N-Acetylneuraminic Acid by Escherichia coli J. Bacteriol., January 1, 1999; 181(1): 47 - 54. [Abstract] [Full Text]

				A.-K. Munster, M. Eckhardt, B. Potvin, M. Muhlenhoff, P. Stanley, and R. Gerardy-Schahn Mammalian cytidine 5'-monophosphate N-acetylneuraminic acid synthetase: A nuclear protein with evolutionarily conserved structural motifs PNAS, August 4, 1998; 95(16): 9140 - 9145. [Abstract] [Full Text] [PDF]

				S. Hinderlich, R. Stasche, R. Zeitler, and W. Reutter 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., September 26, 1997; 272(39): 24313 - 24318. [Abstract] [Full Text] [PDF]

				S. M. Lawrence, K. A. Huddleston, L. R. Pitts, N. Nguyen, Y. C. Lee, W. F. Vann, T. A. Coleman, and M. J. Betenbaugh Cloning and Expression of the Human N-Acetylneuraminic Acid Phosphate Synthase Gene with 2-Keto-3-deoxy-D-glycero- D-galacto-nononic Acid Biosynthetic Ability J. Biol. Chem., June 2, 2000; 275(23): 17869 - 17877. [Abstract] [Full Text] [PDF]

				T. Angata and A. Varki Cloning, Characterization, and Phylogenetic Analysis of Siglec-9, a New Member of the CD33-related Group of Siglecs. EVIDENCE FOR CO-EVOLUTION WITH SIALIC ACID SYNTHESIS PATHWAYS J. Biol. Chem., July 14, 2000; 275(29): 22127 - 22135. [Abstract] [Full Text] [PDF]

				M. Schwarzkopf, K.-P. Knobeloch, E. Rohde, S. Hinderlich, N. Wiechens, L. Lucka, I. Horak, W. Reutter, and R. Horstkorte Sialylation is essential for early development in mice PNAS, April 16, 2002; 99(8): 5267 - 5270. [Abstract] [Full Text] [PDF]