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Information on EC 2.7.7.43 - N-acylneuraminate cytidylyltransferase and Organism(s) Neisseria meningitidis and UniProt Accession P0A0Z8
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2.7.7.43 N-acylneuraminate cytidylyltransferaseIUBMB Comments
Acts on N-acetyl- and N-glycolyl- derivatives.
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Word Map
- 2.7.7.43
-
sialylation
-
sialyltransferase
- meningitidis
-
polysialic
- lipooligosaccharide
- n-acetylmannosamine
- ducreyi
-
sialoglycoconjugates
- synthesis
- kdn
- mannac
- neu5gc
- medicine
- agriculture
The taxonomic range for the selected organisms is: Neisseria meningitidis
The expected taxonomic range for this enzyme is: Eukaryota, Bacteria, Archaea
The expected taxonomic range for this enzyme is: Eukaryota, Bacteria, Archaea
Reaction Schemes
Synonyms
cmp-sialic acid synthetase, cmp-neu5ac synthetase, cmp-neuac synthetase, cmp-neunac synthetase, cmp-n-acetylneuraminic acid synthetase, nmcss, cmp-sia synthetase, dmcss, cmp-sia-syn, dmcsas, 
more
topSYNONYM 
ORGANISM
UNIPROT
COMMENTARY 
LITERATURE
acylneuraminate cytidyltransferase
-
-
-
-
CMP sialate pyrophosphorylase
-
-
-
-
CMP-N-acetylneuraminate synthase
-
-
-
-
CMP-N-acetylneuraminate synthetase
-
-
-
-
CMP-N-acetylneuraminic acid synthase
-
-
-
-
CMP-N-acetylneuraminic acid synthetase
CMP-NANA synthetase
-
-
-
-
CMP-Neu5Ac synthetase
-
-
-
-
CMP-NeuAc synthetase
-
-
-
-
CMP-NeuNAc synthetase
-
-
-
-
CMP-Sia synthetase
-
-
-
-
CMP-sialate synthase
-
-
-
-
CMP-sialate synthetase
-
-
-
-
CMP-sialic acid synthetase
CMP-sialic synthetase
-
-
-
-
CMPsialate pyrophosphorylase
-
-
-
-
CMPsialate synthase
-
-
-
-
cytidine 5'-monophospho-N-acetylneuraminic acid synthetase
-
-
-
-
cytidine 5'-monophosphosialic acid synthetase
-
-
-
-
cytidine 5-monophosphate N-acetylneuraminic acid synthetase
-
-
-
-
cytidine monophosphate-N-acetylneuraminic acid synthetase
-
-
-
-
cytidine monophospho-sialic acid synthetase
-
-
-
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cytidine monophosphoacetylneuraminic synthetase
-
-
-
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cytidine monophosphosialate pyrophosphorylase
-
-
-
-
cytidine monophosphosialate synthetase
-
-
-
-
cytidyltransferase, acylneuraminate
-
-
-
-
cytidylyltransferase, acetylneuraminate
-
-
-
-
sialate cytidylyltransferase
-
-
-
-
CMP-N-acetylneuraminic acid synthetase
-
-
-
-
CMP-sialic acid synthetase
-
-
-
-
REACTION
REACTION DIAGRAM
COMMENTARY
ORGANISM
UNIPROT
LITERATURE
CTP + N-acetylneuraminate = diphosphate + CMP-N-acylneuraminate
typical ordered-sequential kinetic behaviour, enzyme does not bind N-acetylneuraminate in the absence of CTP
REACTION TYPE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
nucleotidyl group transfer
nucleotidyl group transfer
-
-
-
-
PATHWAY SOURCE
PATHWAYS
-
-, -
SYSTEMATIC NAME
IUBMB Comments
CTP:N-acylneuraminate cytidylyltransferase
Acts on N-acetyl- and N-glycolyl- derivatives.
CAS REGISTRY NUMBER
COMMENTARY
9067-82-7
-
SUBSTRATE
PRODUCT
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate)
(Substrate)
LITERATURE
(Substrate)
(Substrate)
COMMENTARY
(Product)
(Product)
LITERATURE
(Product)
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
r=reversible
ir=irreversible
?=not specified
CTP + N-acetylneuraminate
diphosphate + CMP-N-acetylneuraminate
-
-
-
?
CTP + N-glycolylneuraminate
diphosphate + CMP-N-glycolylneuraminate
-
-
-
?
CDP + N-acetylneuraminate
phosphate + CMP-N-acetylneuraminate
-
no activity with CDP
-
-
?
CTP + (2S,4S,5R,6R)-6-((1R,2S)-3-azido-1,2-dihydroxy-propyl)-2,4,5-trihydroxy-6-methyl-tetrahydro-pyran-2-carboxylic acid
diphosphate + CMP-(2S,4S,5R,6R)-6-((1R,2S)-3-azido-1,2-dihydroxy-propyl)-2,4,5-trihydroxy-6-methyl-tetrahydro-pyran-2-carboxylic acid
-
-
-
-
?
CTP + (N-4-O-)diacetylneuraminic acid
diphosphate + CMP-N-4-O-diacetylneuraminate
-
(N-4-O-)diacetylneuraminic acid is 46% effective compared to N-acylneuraminate
-
-
?
CTP + 3-deoxy-D-galacto-2-octulosonic acid
diphosphate + CMP-3-deoxy-D-galacto-2-octulosonic acid
-
-
-
-
?
CTP + 8-O-methyl-N-acetylneuraminate
diphosphate + CMP-(8-O-methyl)-N-acetylneuraminate
-
-
-
-
?
CTP + N-(N-benzyloxycarbonyl-glycyl)-muramic acid
diphosphate + CMP-N-(N-benzyloxycarbonyl-glycyl)-muramic acid
-
-
-
-
?
CTP + N-acetylmuramic acid
diphosphate + CMP-N-acetylmuramic acid
-
-
-
-
?
CTP + N-acetylneuraminate
diphosphate + CMP-N-acetylneuraminate
-
-
-
-
?
CTP + N-azidoacetylmuramic acid
diphosphate + CMP-N-azidoacetylmuramic acid
-
-
-
-
?
CTP + N-azidomuramic acid
diphosphate + CMP-N-azidomuramic acid
-
-
-
-
?
CTP + N-glycolylneuraminate
diphosphate + CMP-N-glycolylneuraminate
-
-
-
-
?
CTP + N-hydroxyacetylmuramic acid
diphosphate + CMP-N-hydroxyacetylmuramic acid
-
-
-
-
?
CTP + N-hydroxymuramic acid
diphosphate + CMP-N-hydroxymuramic acid
-
-
-
-
?
CTP + N-methylglycolylneuraminate
diphosphate + CMP-N-methylglycolylneuraminate
-
-
-
-
?
TTP + N-acylneuraminate
diphosphate + TMP-N-acylneuraminate
-
no activity with TTP
-
-
?
UTP + N-acylneuraminate
diphosphate + UMP-N-acylneuraminate
-
no activity with UTP
-
-
?
CTP + N-acylneuraminate
diphosphate + CMP-N-acylneuraminate
-
-
-
?
CTP + N-acylneuraminate
diphosphate + CMP-N-acylneuraminate
-
-
-
?
CTP + N-acylneuraminate
diphosphate + CMP-N-acylneuraminate
the enzyme is involved in the production of activated sialic acids
-
-
?
CTP + N-acylneuraminate
diphosphate + CMP-N-acylneuraminate
-
-
-
-
?
CTP + N-acylneuraminate
diphosphate + CMP-N-acylneuraminate
-
-
-
-
ir
additional information
?
-
no substrate: 2-keto-3-deoxy-D-glycero-D-galacto-nonulosonic acid, kcat/Km value is about 5000fold lower than that of N-acetylneuraminate
-
-
?
additional information
?
-
-
no substrate: 2-keto-3-deoxy-D-glycero-D-galacto-nonulosonic acid, kcat/Km value is about 5000fold lower than that of N-acetylneuraminate
-
-
?
additional information
?
-
-
the enzyme is used for preparative synthesis of CMP-sialic acid derivatives in a one-pot two-enzyme system with EC 4.1.3.3 and EC 2.7.7.43
-
-
?
additional information
?
-
-
does not accept N-glycolylneuraminic acid and triacetylneuraminic acid as substrates
-
-
?
NATURAL SUBSTRATE
NATURAL PRODUCT
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate)
(Substrate)
LITERATURE
(Substrate)
(Substrate)
COMMENTARY
(Product)
(Product)
LITERATURE
(Product)
(Product)
REVERSIBILITY
r=reversible
ir=irreversible
?=not specified
r=reversible
ir=irreversible
?=not specified
CTP + N-acylneuraminate
diphosphate + CMP-N-acylneuraminate
-
-
-
?
CTP + N-acylneuraminate
diphosphate + CMP-N-acylneuraminate
the enzyme is involved in the production of activated sialic acids
-
-
?
METALS and IONS
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
Mg2+
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
saturated sulfo-UDP ester
-
1 mM, 37°C, pH 8.5, 15 min, 24% inhibition
ACTIVATING COMPOUND
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
additional information
no necessity of including a sulfhydryl reagent, such as dithiothreitol, in the assay reaction
-
additional information
-
no necessity of including a sulfhydryl reagent, such as dithiothreitol, in the assay reaction
-
KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
TURNOVER NUMBER [1/s]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
kcat/KM VALUE [1/mMs-1]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
IC50 VALUE [mM]
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
0.13
2-deoxy-2,3-dehydro-N-acetylneuraminic acid
Neisseria meningitidis
at pH 8.5 and 37°C
additional information
CDP
Neisseria meningitidis
-
1 mM, 37°C, pH 8.5, 15 min, 14% inhibition
additional information
saturated sulfo-CDP
Neisseria meningitidis
-
1 mM, 37°C, pH 8.5, 15 min, 55% inhibition
additional information
saturated sulfo-UDP
Neisseria meningitidis
-
1 mM, 37°C, pH 8.5, 15 min, 35% inhibition
additional information
saturated sulfo-UDP ester
Neisseria meningitidis
-
1 mM, 37°C, pH 8.5, 15 min, 24% inhibition
additional information
sulfo-CDP
Neisseria meningitidis
-
1 mM, 37°C, pH 8.5, 15 min, 47% inhibition
additional information
sulfo-CTP
Neisseria meningitidis
-
1 mM, 37°C, pH 8.5, 15 min, 39% inhibition
additional information
sulfo-UDP
Neisseria meningitidis
-
1 mM, 37°C, pH 8.5, 15 min, 6% inhibition
additional information
sulfo-UTP
Neisseria meningitidis
-
1 mM, 37°C, pH 8.5, 15 min, 33% inhibition
pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
8.5
pH RANGE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
37
ORGANISM
COMMENTARY
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
LOCALIZATION
ORGANISM
UNIPROT
COMMENTARY
GeneOntology No.
LITERATURE
SOURCE
GENERAL INFORMATION
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
physiological function
-
CMP-sialic acid synthetase is a key enzyme in the biosynthesis of the capsular polysaccharides required for bacterial infection
UNIPROT
ENTRY NAME
ORGANISM
NO. OF AA
NO. OF TRANSM. HELICES
MOLECULAR WEIGHT[Da]
SOURCE
SEQUENCE
LOCALIZATION PREDICTION
The reliability class of the localization prediction ranges from 1 (very reliable) to 5 (not reliable).
The reliability class of the localization prediction ranges from 1 (very reliable) to 5 (not reliable). PDB
SCOP
CATH
UNIPROT
ORGANISM
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
SUBUNIT
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
CRYSTALLIZATION (Commentary)
ORGANISM
UNIPROT
LITERATURE
ligand-free and sialic acid-bound enzyme, sitting drop vapor diffusion method, using 0.16 M calcium acetate, 0.08 M sodium cacodylate pH 6.5, 14.4% (w/v) PEG 8000, and 20% (v/v) glycerol. Enzyme in complex with Ca2+ and CTP, sitting drop vapor diffusion method, using 0.1 M imidazole pH 8.0, 6% (w/v) PEG 8000, and 0.2 M calcium acetate
modeling of the closed conformation. Enzyme forms a dimer complex. Amino acids involved in the Neu5Ac binding pocket are Glu162 and Arg165 from monomer B and Gln104, Ser82, Val86, Tyr179, and Phe192 from monomer A
-
PROTEIN VARIANTS
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
D209A
D211A
E162A
the mutant enzyme shows 14.5% activity compared to the wild type enzyme
E162Q
the mutant enzyme shows 15.3% activity compared to the wild type enzyme
K142A
N175A
16fold increases in the Km value for CTP and 23fold for N-acetylneuraminate
Q104E
dramatic loss of activity. Residue involved in metal binding
Q104L
dramatic loss of activity. Residue involved in metal binding
Q104N
dramatic loss of activity. Residue involved in metal binding
R165A
the mutant enzyme shows 0.163% activity compared to the wild type enzyme
S31R
-
mutant displays improved substrate promiscuity, catalytic activities for substrates N-glycolylneuraminate, N-methylglycolylneuraminate and 8-O-methyl N-acetyl-neuraminate are improved compared to wild-type
D209A
almost complete loss of activity. Residue involved in metal binding
D211A
dramatic loss of activity. Residue involved in metal binding
K142A
10000fold reduction in kcat with an insignificant, fourfold, rise in Km for CTP
K142A
dramatic loss of activity. Residue involved in metal binding
STORAGE STABILITY
ORGANISM
UNIPROT
LITERATURE
4°C, 7 months, a magnetic nanoparticle-CSS preparation retains almost 100% activity
-
PURIFICATION (Commentary)
ORGANISM
UNIPROT
LITERATURE
CLONED (Commentary)
ORGANISM
UNIPROT
LITERATURE
the enzyme is subcloned for overexpression in Escherichia coli K12 using the expression vector pKK233-3
-
transformation of the CMP-NeuNAc defective Escherichia coli K1 strain EV5 with CMP-NeuNAc synthetase from Neisseria meningitidis can complement the defect in Escherichia coli
-
APPLICATION
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
medicine
the enzyme is involved in the production of activated sialic acids. Sialic acids are virulence factors in a variety of bacterial species, e.g. Neisseria menigitidis. As such, the identification of the bacterial CMP-NeuAc synthetase active site can serve as a starting point for rational drug design strategies
synthesis
since CMP-N-acylneuraminate is unstable and relatively expensive, the enzyme is valuable for the preparative enzymatic synthesis of silylated oligosaccharides
synthesis
synthesis
-
the high expressivity of the recombinant production clone, the high catalytic efficiency of the enzyme, and its broad substrate tolerance make this enzyme the preferred catalyst for the enzymatic synthesis of CMP-Neu5Ac
synthesis
-
the enzyme is used for preparative synthesis of CMP-sialic acid derivatives in a one-pot two-enzyme system with EC 4.1.3.3 and EC 2.7.7.43
REF.
AUTHORS
TITLE
JOURNAL
VOL.
PAGES
YEAR
ORGANISM (UNIPROT)
PUBMED ID
SOURCE
Edwards, U.; Frosch, M.
Sequence and functional analysis of the cloned Neisseria meningitidis CMP neunac synthetase
FEMS Microbiol. Lett.
96
161-166
1992
Neisseria meningitidis
-
Karwaski, M.F.; Wakarchuk, W.W.; Gilbert, M.
High-level expression of recombinant Neisseria CMP-sialic acid synthetase in Escherichia coli
Protein Expr. Purif.
25
237-240
2002
Neisseria meningitidis (P0A0Z8), Neisseria meningitidis, Neisseria meningitidis 406Y (P0A0Z8)
Gilbert, M.; Watson, D.C.; Wakarchuk, W.W.
Purification and characterization of the recombinant CMP-sialic acid synthetase from Neisseria meningitidis
Biotechnol. Lett.
19
417-420
1997
Neisseria meningitidis (P0A0Z8), Neisseria meningitidis 406Y (P0A0Z8)
-
Mosimann, S.C.; Gilbert, M.; Dombroswki, D.; To, R.; Wakarchuk, W.; Strynadka, N.C.
Structure of a sialic acid-activating synthetase, CMP-acylneuraminate synthetase in the presence and absence of CDP
J. Biol. Chem.
276
8190-8196
2001
Neisseria meningitidis (P0A0Z8), Neisseria meningitidis
Knorst, M.; Fessner, W.D.
CMP-sialate synthetase from Neisseria meningitidis - overexpression and application to the synthesis of oligosaccharides containing modified sialic acids
Adv. Synth. Catal.
343
698-710
2001
Neisseria meningitidis
-
Bravo, I.G.; Reglero, A.
The cytidylyltransferases family: properties, kinetics, genomic and phylogeny: The cytidylyltransferases family: properties, kinetics, genomic and phylogeny
Recent Res. Devel. Biochem.
4
223-254
2003
Bos taurus, Escherichia coli, Equus caballus, [Haemophilus] ducreyi, Cricetinae, Erinaceidae, Homo sapiens, Mannheimia haemolytica, Neisseria meningitidis, Oncorhynchus mykiss, Rattus norvegicus, Streptococcus agalactiae, Sus scrofa
-
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.
12
6427-6435
2004
Neisseria meningitidis
Mizanur, R.M.; Pohl, N.L.
Bacterial CMP-sialic acid synthetases: production, properties, and applications
Appl. Microbiol. Biotechnol.
80
757-765
2008
Acetivibrio thermocellus, Escherichia coli, [Haemophilus] ducreyi, Mannheimia haemolytica, Neisseria meningitidis, Escherichia coli K-235, Mannheimia haemolytica A2
Yu, C.C.; Lin, P.C.; Lin, C.C.
Site-specific immobilization of CMP-sialic acid synthetase on magnetic nanoparticles and its use in the synthesis of CMP-sialic acid
Chem. Commun. (Camb. )
21
1308-1310
2008
Neisseria meningitidis
Wong, J.H.; 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.
7
27-29
2009
Neisseria meningitidis
Li, Y.; Yu, H.; Cao, H.; Muthana, S.; Chen, X.
Pasteurella multocida CMP-sialic acid synthetase and mutants of Neisseria meningitidis CMP-sialic acid synthetase with improved substrate promiscuity
Appl. Microbiol. Biotechnol.
93
2411-2423
2012
[Haemophilus] ducreyi, Neisseria meningitidis, Pasteurella multocida, Pasteurella multocida ATCC 15742
Horsfall, L.E.; Nelson, A.; Berry, A.
Identification and characterization of important residues in the catalytic mechanism of CMP-Neu5Ac synthetase from Neisseria meningitidis
FEBS J.
277
2779-2790
2010
Neisseria meningitidis (P0A0Z8), Neisseria meningitidis
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