n acylneuraminate cytidylyltransferase


Summary: An enzyme that forms CMP-acylneuraminic acids, which donate the N-acylneuraminic acid residues to the terminal sugar residue of a ganglioside or glycoprotein. EC

Top Publications

  1. Zapata G, Vann W, Aaronson W, Lewis M, Moos M. Sequence of the cloned Escherichia coli K1 CMP-N-acetylneuraminic acid synthetase gene. J Biol Chem. 1989;264:14769-74 pubmed
    ..Gel filtration and ultracentrifugation experiments under nondenaturing conditions suggest that the enzyme is active as a 49,000-dalton monomer but may form aggregates. ..
  2. Tullius M, Munson R, Wang J, Gibson B. Purification, cloning, and expression of a cytidine 5'-monophosphate N-acetylneuraminic acid synthetase from Haemophilus ducreyi. J Biol Chem. 1996;271:15373-80 pubmed
    ..In contrast to other bacterial CMP-NeuAc synthetases, the H. ducreyi enzyme exhibited a different substrate specificity, being capable of also using N-glycolylneuraminic acid as a substrate. ..
  3. Haft R, Wessels M. Characterization of CMP-N-acetylneuraminic acid synthetase of group B streptococci. J Bacteriol. 1994;176:7372-4 pubmed
    ..4 for CTP and a pH optimum of 8.3 to 9.4, required magnesium, and was stimulated by dithiothreitol. This is the first characterization of an enzyme involved in group B streptococcus capsular polysaccharide biosynthesis. ..
  4. Munster A, Weinhold B, Gotza B, Muhlenhoff M, Frosch M, Gerardy Schahn R. Nuclear localization signal of murine CMP-Neu5Ac synthetase includes residues required for both nuclear targeting and enzymatic activity. J Biol Chem. 2002;277:19688-96 pubmed
    ..Combined, the data demonstrate that nuclear transport and enzyme activity are independent functions that share some common amino acid requirements in CMP-Neu5Ac synthetase. ..
  5. Jelakovic S, Schulz G. The structure of CMP:2-keto-3-deoxy-manno-octonic acid synthetase and of its complexes with substrates and substrate analogs. J Mol Biol. 2001;312:143-55 pubmed
    ..The N-terminal domain shows sequence homology with the CMP-NeuAc synthetases. Moreover, the chain fold and the substrate-binding position of CKS resemble those of other enzymes processing nucleotide-sugars. ..
  6. Vann W, Silver R, Abeijon C, Chang K, Aaronson W, Sutton A, et al. Purification, properties, and genetic location of Escherichia coli cytidine 5'-monophosphate N-acetylneuraminic acid synthetase. J Biol Chem. 1987;262:17556-62 pubmed
    ..Both the enzyme and the 50,000 Mr polypeptide have the same NH2-terminal amino acid sequence. Antibodies prepared to a peptide derived from the NH2-terminal amino acid sequence bind to purified CMP-NeuAc synthetase. ..
  7. Tullius M, Vann W, Gibson B. Covalent modification of Lys19 in the CTP binding site of cytidine 5'-monophosphate N-acetylneuraminic acid synthetase. Protein Sci. 1999;8:666-75 pubmed
    ..Lys19, a residue conserved in CMP-NeuAc synthetases, was identified as being labeled with the beta-elimination product of oCTP. ..
  8. Nakata D, M nster A, Gerardy Schahn R, Aoki N, Matsuda T, Kitajima K. Molecular cloning of a unique CMP-sialic acid synthetase that effectively utilizes both deaminoneuraminic acid (KDN) and N-acetylneuraminic acid (Neu5Ac) as substrates. Glycobiology. 2001;11:685-92 pubmed
    ..These results indicate that at least one cloned enzyme has the ability to utilize both KDN and Neu5Ac as substrates efficiently and is useful for the production of CMP-KDN...
  9. Samuels N, Gibson B, Miller S. Investigation of the kinetic mechanism of cytidine 5'-monophosphate N-acetylneuraminic acid synthetase from Haemophilus ducreyi with new insights on rate-limiting steps from product inhibition analysis. Biochemistry. 1999;38:6195-203 pubmed
    ..To our knowledge, this relationship has not been previously recognized. ..

More Information


  1. Mizanur R, Pohl N. Bacterial CMP-sialic acid synthetases: production, properties, and applications. Appl Microbiol Biotechnol. 2008;80:757-65 pubmed publisher
    ..The CSSs are important enzymes for the chemoenzymatic synthesis of various sialooligosaccharides of significance in biotechnology. ..
  2. Mosimann S, Gilbert M, Dombroswki D, To R, Wakarchuk W, Strynadka N. Structure of a sialic acid-activating synthetase, CMP-acylneuraminate synthetase in the presence and absence of CDP. J Biol Chem. 2001;276:8190-6 pubmed publisher
    ..g. Neisseria, Haemophilus, group B streptococci, etc.). As such, the identification of the bacterial CMP-NeuAc synthetase active site can serve as a starting point for rational drug design strategies...
  3. Ambrose M, Freese S, Reinhold M, Warner T, Vann W. 13C NMR investigation of the anomeric specificity of CMP-N-acetylneuraminic acid synthetase from Escherichia coli. Biochemistry. 1992;31:775-80 pubmed
    ..Thus, CMP-beta-NeuAc is probably formed by a direct transfer of the anomeric oxygen of beta-NeuAc to the alpha-phosphate of CTP. ..
  4. Kean E. Sialic acid activation. Glycobiology. 1991;1:441-7 pubmed
    ..This report does not deal with its utilization by the sialyltransferases. ..
  5. Krapp S, Münster Kühnel A, Kaiser J, Huber R, Tiralongo J, Gerardy Schahn R, et al. The crystal structure of murine CMP-5-N-acetylneuraminic acid synthetase. J Mol Biol. 2003;334:625-37 pubmed
    ..In addition, a complex regulation of the enzyme activity is proposed that includes phosphorylation and dephosphorylation. ..
  6. Münster A, Eckhardt M, Potvin B, Muhlenhoff M, Stanley P, Gerardy Schahn R. Mammalian cytidine 5'-monophosphate N-acetylneuraminic acid synthetase: a nuclear protein with evolutionarily conserved structural motifs. Proc Natl Acad Sci U S A. 1998;95:9140-5 pubmed
    ..Four stretches of amino acids that occur in the N-terminal region are highly conserved in bacterial CMP-Neu5Ac synthetases, providing evidence for an ancestral relationship between the sialylation pathways of bacterial and animal cells. ..
  7. Stoughton D, Zapata G, Picone R, Vann W. Identification of Arg-12 in the active site of Escherichia coli K1 CMP-sialic acid synthetase. Biochem J. 1999;343 Pt 2:397-402 pubmed
    ..Steady-state kinetic analysis reveals changes in k(cat), K(m) and K(s) for CTP, which implicates Arg-12 in catalysis and substrate binding. ..
  8. Lin C, Lin C. Enzymatic and chemical approaches for the synthesis of sialyl glycoconjugates. Adv Exp Med Biol. 2001;491:215-30 pubmed
    ..Nevertheless, reducing the number of reaction steps (for preparing both donors and acceptors of glycosylation), and enhancing stereoselectivity, as well as reaction yield are still problems to be overcome. ..
  9. Stangier P, Treder W, Thiem J. Chemoenzymatic galactosialylation with integrated cofactor regeneration. Glycoconj J. 1993;10:26-33 pubmed
  10. Bravo I, Garcia Vallve S, Romeu A, Reglero A. Prokaryotic origin of cytidylyltransferases and alpha-ketoacid synthases. Trends Microbiol. 2004;12:120-8 pubmed
  11. Edwards U, Frosch M. Sequence and functional analysis of the cloned Neisseria meningitidis CMP-NeuNAc synthetase. FEMS Microbiol Lett. 1992;75:161-6 pubmed
    ..coli K1. Enzymatic activity was confirmed in vitro and in vivo. Transformation of the CMP-NeuNAc defective E. coli K1 strain EV5 with the meningococcal CMP-NeuNAc synthetase could complement the defect in E. coli...
  12. Humphrey A, Fremann C, Critchley P, Malykh Y, Schauer R, Bugg T. Biological properties of N-acyl and N-haloacetyl neuraminic acids: processing by enzymes of sialic acid metabolism, and interaction with influenza virus. Bioorg Med Chem. 2002;10:3175-85 pubmed
    ..CMP-N-propionyl neuraminic acid was found not to be a substrate for CMP-N-acetyl neuraminic acid hydroxylase from pig submandibular gland. ..
  13. Yu H, Ryan W, Yu H, Chen X. Characterization of a bifunctional cytidine 5'-monophosphate N-acetylneuraminic acid synthetase cloned from Streptococcus agalactiae. Biotechnol Lett. 2006;28:107-13 pubmed
    ..0. A metal cofactor (either Mg(2+) or Mn(2+)) is required for the CMP-sialic acid synthetase activity but is not for acetylhydrolase activity. Both catalytic functions, however, are impaired by high concentrations of Mn(2+). ..
  14. Barrallo S, Reglero A, Revilla Nuin B, Martínez Blanco H, Rodriguez Aparicio L, Ferrero M. Regulation of capsular polysialic acid biosynthesis by temperature in Pasteurella haemolytica A2. FEBS Lett. 1999;445:325-8 pubmed
    ..These findings indicate that the cellular growth temperature regulates the synthesis of these enzymes and hence the concentration of the intermediates necessary for capsular polysaccharide genesis in P. haemolytica A2. ..
  15. Haselhorst T, Münster Kühnel A, Oschlies M, Tiralongo J, Gerardy Schahn R, von Itzstein M. Direct detection of ligand binding to Sepharose-immobilised protein using saturation transfer double difference (STDD) NMR spectroscopy. Biochem Biophys Res Commun. 2007;359:866-70 pubmed
  16. Ikeda K, Nagao Y, Achiwa K. Synthesis of sialic acid-containing nucleotide sugars: CMP-sialic acid analogs. Carbohydr Res. 1992;224:123-31 pubmed
  17. Jin C, Jin C. [Minimal functional domain of cytidine 5'-monophosphate N-acetylneuraminic acid (CMP-NeuAc) synthetase from Escherichia coli]. Sheng Wu Gong Cheng Xue Bao. 2002;18:676-82 pubmed
    ..coli CMP-NeuAc synthetase could affect the conformation of the enzymatic catalytic domain and therefore affect its catalytic activity and thermostability, although it is not involved in enzymatic activity directly. ..
  18. Carette J, Guimaraes C, Varadarajan M, Park A, Wuethrich I, Godarova A, et al. Haploid genetic screens in human cells identify host factors used by pathogens. Science. 2009;326:1231-5 pubmed publisher
    ..This approach has both conceptual and practical parallels with genetic approaches in haploid yeast. ..
  19. Liu G, Jin C, Jin C. CMP-N-acetylneuraminic acid synthetase from Escherichia coli K1 is a bifunctional enzyme: identification of minimal catalytic domain for synthetase activity and novel functional domain for platelet-activating factor acetylhydrolase activity. J Biol Chem. 2004;279:17738-49 pubmed
    ..Although its physiological function is still unclear, it has been proposed that the alpha1-subunit-like domain of E. coli may be involved in the traversal of the blood-brain barrier. ..
  20. Lawrence S, Huddleston K, Tomiya N, Nguyen N, Lee Y, Vann W, et al. Cloning and expression of human sialic acid pathway genes to generate CMP-sialic acids in insect cells. Glycoconj J. 2001;18:205-13 pubmed
    ..Engineering the CMP-SA metabolic pathway may be beneficial in various cell lines in which CMP-Neu5Ac production limits sialylation of glycoproteins or other glycans. ..
  21. Ratzow S, Gaia V, Helbig J, Fry N, Lück P. Addition of neuA, the gene encoding N-acylneuraminate cytidylyl transferase, increases the discriminatory ability of the consensus sequence-based scheme for typing Legionella pneumophila serogroup 1 strains. J Clin Microbiol. 2007;45:1965-8 pubmed
    ..932 to 0.963) and subdivided some known large common complexes (e.g., 1,4,3,1,1,1). This modification to the standard method is proposed as the method of choice in the epidemiological investigation of L. pneumophila infections. ..
  22. Gross H, Kovac A, Rose U, Watzlawick H, Brossmer R. Interaction of N-acetyl-4-epi-D-neuraminic acid with key enzymes of sialic acid metabolism. Biochemistry. 1988;27:4279-83 pubmed
    ..Therefore, no conclusions about the mechanism of the inhibition could be reached. This is the first reported sialic acid analogue which can act as an inhibitor of CMP-sialic acid synthase. ..
  23. Hood D, Makepeace K, Deadman M, Rest R, Thibault P, Martin A, et al. Sialic acid in the lipopolysaccharide of Haemophilus influenzae: strain distribution, influence on serum resistance and structural characterization. Mol Microbiol. 1999;33:679-92 pubmed
    ..We determine for the first time the site of attachment of sialic acid in the LPS of a non-typeable strain and report that a small proportion of glycoforms include two sialic acid residues in a disaccharide unit. ..
  24. Hartlieb S, Günzel A, Gerardy Schahn R, Münster Kühnel A, Kirschning A, Dräger G. Chemoenzymatic synthesis of CMP-N-acetyl-7-fluoro-7-deoxy-neuraminic acid. Carbohydr Res. 2008;343:2075-82 pubmed publisher
    ..Finally, conditions were optimized to perform a one-pot reaction starting from fluorinated mannosamine, which yielded the 7-fluoro-7-deoxy-CMP-sialic acid by incubation with three enzymes. ..
  25. Martin R, Witte K, Wong C. The synthesis and enzymatic incorporation of sialic acid derivatives for use as tools to study the structure, activity, and inhibition of glycoproteins and other glycoconjugates. Bioorg Med Chem. 1998;6:1283-92 pubmed
    ..Compound 3 was enzymatically incorporated into an oligosaccharide and a non-natural glycoprotein for use as an aid in the structure determination of these compounds by X-ray crystallography. ..
  26. Viswanathan K, Tomiya N, Park J, Singh S, Lee Y, Palter K, et al. Expression of a functional Drosophila melanogaster CMP-sialic acid synthetase. Differential localization of the Drosophila and human enzymes. J Biol Chem. 2006;281:15929-40 pubmed
  27. Lubineau A, Auge C, Gautheron Le Narvor C, Ginet J. Combined chemical and enzymatic synthesis of the sialylated non reducing terminal sequence of GM1b glycolylated ganglioside, a potential human tumor marker. Bioorg Med Chem. 1994;2:669-74 pubmed
    ..alpha-Neu5Gc-(2-3)-beta-D-Gal-(1-3)-beta-D-GalNac-OBn 13 isolated in 56% yield was deprotected to give the non-reducing terminal sequence of GM1b glycolylated ganglioside, which might be expressed in human tumors. ..
  28. Tiralongo J, Fujita A, Sato C, Kitajima K, Lehmann F, Oschlies M, et al. The rainbow trout CMP-sialic acid synthetase utilises a nuclear localization signal different from that identified in the mouse enzyme. Glycobiology. 2007;17:945-54 pubmed
  29. González Clemente C, Luengo J, Rodriguez Aparicio L, Ferrero M, Reglero A. High production of polysialic acid [Neu5Ac alpha(2-8)-Neu5Ac alpha(2-9)]n by Escherichia coli K92 grown in a chemically defined medium. Regulation of temperature. Biol Chem Hoppe Seyler. 1990;371:1101-6 pubmed
    ..The enzyme, CMP-Neu5Ac synthetase, was not detected in cells grown at 20 degrees C. The lack of this enzyme explains the absence of polymer biosynthesis when the bacterium was grown at 20 degrees C. ..
  30. Christian R, Schreiner E, Zbiral E, Schulz G. The side-chain conformations of N-acetyl-7-,8-,9-deoxy-, and -4,7-dideoxy-neuraminic acid and their effect on the activation of CTP:N-acylneuraminic acid cytidylyltransferase. Carbohydr Res. 1989;194:49-61 pubmed
    ..The conformations were correlated with the activity with CTP: N-acetylneuraminic acid cytidylyltransferase (CMPsialate synthase; EC ..
  31. Unverzagt C, Kelm S, Paulson J. Chemical and enzymatic synthesis of multivalent sialoglycopeptides. Carbohydr Res. 1994;251:285-301 pubmed
    ..A possible mechanism for increased binding is proposed. ..
  32. Hamamoto T, Takeda S, Noguchi T. Enzymatic synthesis of cytidine 5'-monophospho-N-acetylneuraminic acid. Biosci Biotechnol Biochem. 2005;69:1944-50 pubmed
    ..coli cells, and H. influenzae CMP-NeuAc synthetase, and succeeded in efficient production of CMP-NeuAc due to a sufficient supply of PEP and efficient conversion of CMP to cytidine 5'-triphosphate by yeast cells. ..
  33. Yu H, Yu H, Karpel R, Chen X. Chemoenzymatic synthesis of CMP-sialic acid derivatives by a one-pot two-enzyme system: comparison of substrate flexibility of three microbial CMP-sialic acid synthetases. Bioorg Med Chem. 2004;12:6427-35 pubmed
    ..meningitidis synthetase and the aldolase. ..
  34. Rodriguez Aparicio L, Luengo J, González Clemente C, Reglero A. Purification and characterization of the nuclear cytidine 5'-monophosphate N-acetylneuraminic acid synthetase from rat liver. J Biol Chem. 1992;267:9257-63 pubmed
    ..6 mM), whereas CMP-NeuAc, high CTP concentrations, and other nucleotides (CDP, CMP, ATP, UTP, GTP, and TTP) inhibited the enzyme to different extents. ..
  35. Cao H, Muthana S, Li Y, Cheng J, Chen X. Parallel chemoenzymatic synthesis of sialosides containing a C5-diversified sialic acid. Bioorg Med Chem Lett. 2009;19:5869-71 pubmed publisher
  36. Rauvolfova J, Venot A, Boons G. Chemo-enzymatic synthesis of C-9 acetylated sialosides. Carbohydr Res. 2008;343:1605-11 pubmed publisher
    ..The N(alpha)-CBz-protected intermediate 11 can be employed for the synthesis of glycolipopeptides for immunization purposes. ..
  37. Viswanathan K, Narang S, Hinderlich S, Lee Y, Betenbaugh M. Engineering intracellular CMP-sialic acid metabolism into insect cells and methods to enhance its generation. Biochemistry. 2005;44:7526-34 pubmed
  38. Kean E, Münster Kühnel A, Gerardy Schahn R. CMP-sialic acid synthetase of the nucleus. Biochim Biophys Acta. 2004;1673:56-65 pubmed
    ..In addition, we take the advantage to discuss most recent developments towards understanding structure--function relations of this enzyme. ..
  39. Münster Kühnel A, Tiralongo J, Krapp S, Weinhold B, Ritz Sedlacek V, Jacob U, et al. Structure and function of vertebrate CMP-sialic acid synthetases. Glycobiology. 2004;14:43R-51R pubmed
  40. Ganguli S, Zapata G, Wallis T, Reid C, Boulnois G, Vann W, et al. Molecular cloning and analysis of genes for sialic acid synthesis in Neisseria meningitidis group B and purification of the meningococcal CMP-NeuNAc synthetase enzyme. J Bacteriol. 1994;176:4583-9 pubmed
    ..coli K1, two proteins involved in the synthesis of NeuNAc. These results indicate that common steps exist in the biosynthesis of NeuNAc in these two microorganisms...
  41. Lee S, Lee J, Yi J, Kim B. Production of cytidine 5'-monophosphate N-acetylneuraminic acid using recombinant Escherichia coli as a biocatalyst. Biotechnol Bioeng. 2002;80:516-24 pubmed
    ..coli extract and alpha2,3-sialyltransferase. ..
  42. Bozue J, Tullius M, Wang J, Gibson B, Munson R. Haemophilus ducreyi produces a novel sialyltransferase. Identification of the sialyltransferase gene and construction of mutants deficient in the production of the sialic acid-containing glycoform of the lipooligosaccharide. J Biol Chem. 1999;274:4106-14 pubmed
    ..We conclude that the lst gene encodes the H. ducreyi sialyltransferase. Since the lst gene product has little, if any, structural relationship to other sialyltransferases, this protein represents a new class of sialyltransferase...
  43. Ishige K, Hamamoto T, Shiba T, Noguchi T. Novel method for enzymatic synthesis of CMP-NeuAc. Biosci Biotechnol Biochem. 2001;65:1736-40 pubmed
  44. Misaki R, Fujiyama K, Seki T. Expression of human CMP-N-acetylneuraminic acid synthetase and CMP-sialic acid transporter in tobacco suspension-cultured cell. Biochem Biophys Res Commun. 2006;339:1184-9 pubmed
    ..The recombinant human proteins showed their biological activities. These results show that the plant cell can be a useful bioreactor for the production of mammalian glycoproteins. ..
  45. Antoine T, Priem B, Heyraud A, Greffe L, Gilbert M, Wakarchuk W, et al. Large-scale in vivo synthesis of the carbohydrate moieties of gangliosides GM1 and GM2 by metabolically engineered Escherichia coli. Chembiochem. 2003;4:406-12 pubmed
    ..In high-cell-density cultures, the production yields for the GM2 and GM1 oligosaccharides were 1.25 g L(-1) and 0.89 g L(-1), respectively. ..
  46. Haft R, Wessels M, Mebane M, Conaty N, Rubens C. Characterization of cpsF and its product CMP-N-acetylneuraminic acid synthetase, a group B streptococcal enzyme that can function in K1 capsular polysaccharide biosynthesis in Escherichia coli. Mol Microbiol. 1996;19:555-63 pubmed
    ..We concluded that cpsF encodes CMP-Neu5Ac synthetase in type III GBS, and that the GBS enzyme can function in the capsule-synthesis of a heterologous bacterial species...
  47. Shames S, Simon E, Christopher C, Schmid W, Whitesides G, Yang L. CMP-N-acetylneuraminic acid synthetase of Escherichia coli: high level expression, purification and use in the enzymatic synthesis of CMP-N-acetylneuraminic acid and CMP-neuraminic acid derivatives. Glycobiology. 1991;1:187-91 pubmed
    ..coli synthetase, together with the high degree of purity readily obtainable from crude cell extracts, make the recombinant bacterial enzyme the preferred catalyst for the enzymatic synthesis of CMP-N-acetylneuraminic acid. ..
  48. Revilla Nuin B, Reglero A, Feo J, Rodriguez Aparicio L, Ferrero M. Identification, expression and tissue distribution of cytidine 5'-monophosphate N-acetylneuraminic acid synthetase activity in the rat. Glycoconj J. 1998;15:233-41 pubmed
    ..Changes in the level of sialylation during development were also found to be intimately related to variations in the expression of this enzyme, at least in brain, heart, kidney, stomach, intestine and lung. ..
  49. Fierfort N, Samain E. Genetic engineering of Escherichia coli for the economical production of sialylated oligosaccharides. J Biotechnol. 2008;134:261-5 pubmed publisher
    ..This high concentration and low cost of fermentation medium should make possible to use sialylated oligosaccharides in new fields such as the food industry...
  50. Zapata G, Crowley J, Vann W. Sequence and expression of the Escherichia coli K1 neuC gene product. J Bacteriol. 1992;174:315-9 pubmed
    ..Both the neuA and neuC genes are coexpressed under the control of a single upstream T7 or tac promoter, suggesting that neuA and neuC are part of an operon. ..
  51. Wong J, Sahni U, Li Y, Chen X, Gervay Hague J. Synthesis of sulfone-based nucleotide isosteres: identification of CMP-sialic acid synthetase inhibitors. Org Biomol Chem. 2009;7:27-9 pubmed publisher
    ..These analogs were shown to be inhibitors of Neisseria meningitidis CSS (NmCSS), which is a key enzyme in the biosynthesis of the capsular polysaccharides required for bacterial infection. ..
  52. Puente Polledo L, Reglero A, González Clemente C, Rodriguez Aparicio L, Ferrero M. Biochemical conditions for the production of polysialic acid by Pasteurella haemolytica A2. Glycoconj J. 1998;15:855-61 pubmed
    ..5 mM, respectively, and finally a membrane-associated polysialyltransferase, which catalyzes the incorporation of sialic acid from CMP-Neu5Ac into polymeric products with an apparent CMP-Neu5Ac Km of 250 microM. ..
  53. Vionnet J, Concepcion N, Warner T, Zapata G, Hanover J, Vann W. Purification of CMP-N-acetylneuraminic acid synthetase from bovine anterior pituitary glands. Glycobiology. 1999;9:481-7 pubmed
    ..The active enzyme chromatographed on a gel filtration column at 158 kDa suggesting it exists in its native form as an oligomer. ..