Information on EC 2.7.7.43 - N-acylneuraminate cytidylyltransferase

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The expected taxonomic range for this enzyme is: Eukaryota, Bacteria

EC NUMBER
COMMENTARY
2.7.7.43
-
RECOMMENDED NAME
GeneOntology No.
N-acylneuraminate cytidylyltransferase
REACTION
REACTION DIAGRAM
COMMENTARY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
CTP + N-acetylneuraminate = diphosphate + CMP-N-acylneuraminate
show the reaction diagram
typical ordered-sequential kinetic behaviour, enzyme does not bind N-acetylneuraminate in the absence of CTP
-
CTP + N-acetylneuraminate = diphosphate + CMP-N-acylneuraminate
show the reaction diagram
-
-
-
-
CTP + N-acylneuraminate = diphosphate + CMP-N-acylneuraminate
show the reaction diagram
ping pong bi bi mechanism with CTP as the first substrate and CMP-NeuAc as the last product
-
CTP + N-acylneuraminate = diphosphate + CMP-N-acylneuraminate
show the reaction diagram
ordered bi-bi kinetic mechanism. CTP binds first and CMP-NeuAc dissociates last. The rate-limiting step appears to be dissociation of CMP-NeuAc
-
CTP + N-acylneuraminate = diphosphate + CMP-N-acylneuraminate
show the reaction diagram
ping pong bi bi mechanism with CTP as the first substrate and CMP-NeuAc as the last product
Mannheimia haemolytica A2
-
-
CTP + N-acylneuraminate = diphosphate + CMP-N-acylneuraminate
show the reaction diagram
-
-
-
-
REACTION TYPE
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
nucleotidyl group transfer
-
-
-
-
nucleotidyl group transfer
-
-
nucleotidyl group transfer
-
-
PATHWAY
KEGG Link
MetaCyc Link
Amino sugar and nucleotide sugar metabolism
-
Biosynthesis of secondary metabolites
-
CMP-N-acetylneuraminate biosynthesis I (eukaryotes)
-
CMP-N-acetylneuraminate biosynthesis II (bacteria)
-
Metabolic pathways
-
SYSTEMATIC NAME
IUBMB Comments
CTP:N-acylneuraminate cytidylyltransferase
Acts on N-acetyl- and N-glycolyl- derivatives.
SYNONYMS
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
acylneuraminate cytidyltransferase
-
-
-
-
Cmas1
Q0E671
-
Cmas2
H9BFW7
-
CMP sialate pyrophosphorylase
-
-
-
-
CMP-N-acetylneuraminate synthase
-
-
-
-
CMP-N-acetylneuraminate synthetase
-
-
-
-
CMP-N-acetylneuraminic acid synthase
-
-
-
-
CMP-N-acetylneuraminic acid synthetase
-
-
-
-
CMP-N-acetylneuraminic acid synthetase
-
-
CMP-N-acetylneuraminic acid synthetase
-
-
CMP-N-acetylneuraminic acid synthetase
Escherichia coli K-235, Escherichia coli K1
-
-
-
CMP-N-acetylneuraminic acid synthetase
-
-
CMP-N-acetylneuraminic acid synthetase
-
-
CMP-N-acetylneuraminic acid synthetase
-
-
CMP-N-acetylneuraminic acid synthetase
Mannheimia haemolytica A2
-
-
-
CMP-N-acetylneuraminic acid synthetase
-
-
CMP-N-acylneuraminic acid synthetase
-
-
CMP-N-acylneuraminic acid synthetase
Clostridium thermocellum ATCC27405
-
-
-
CMP-NANA synthetase
-
-
-
-
CMP-Neu5Ac synthetase
-
-
-
-
CMP-Neu5Ac synthetase
P0A0Z8
-
CMP-Neu5Ac synthetase
-
-
CMP-NeuAc synthetase
-
-
-
-
CMP-NeuAc synthetase
-
-
CMP-NeuAc synthetase
Clostridium thermocellum ATCC27405
-
-
-
CMP-NeuAc synthetase
-
-
CMP-NeuAc synthetase
-
-
-
CMP-NeuNAc synthetase
-
-
-
-
CMP-Sia synthetase
-
-
-
-
CMP-Sia synthetase
-
-
CMP-Sia-syn
Q90WG6
-
CMP-sialate synthase
-
-
-
-
CMP-sialate synthetase
-
-
-
-
CMP-sialic acid synthetase
-
-
-
-
CMP-sialic acid synthetase
-
-
CMP-sialic acid synthetase
-
-
CMP-sialic acid synthetase
-
-
CMP-sialic acid synthetase
Q99KK2
-
CMP-sialic acid synthetase
-
-
CMP-sialic acid synthetase
Q90WG6
-
CMP-sialic acid synthetase (CSS)
-
-
CMP-sialic synthetase
-
-
-
-
CMPsialate pyrophosphorylase
-
-
-
-
CMPsialate synthase
-
-
-
-
CSS
Clostridium thermocellum ATCC27405
-
-
-
CSS
Escherichia coli K-235
-
-
-
CSS
Mannheimia haemolytica A2
-
-
-
CSS
Q99KK2
-
CSS
Pasteurella multocida ATCC 15742
-
-
-
cytidine 5'-monophosphate N-acetylneuraminic acid synthetase
-
-
cytidine 5'-monophosphate N-acetylneuraminic acid synthetase
Escherichia coli K-235
-
-
-
cytidine 5'-monophosphate N-acetylneuraminic acid synthetase
-
-
cytidine 5'-monophosphate N-acetylneuraminic acid synthetase
Mannheimia haemolytica A2
-
-
-
cytidine 5'-monophosphate N-acetylneuraminic acid synthetase
-
-
cytidine 5'-monophospho-N-acetylneuraminic acid synthetase
-
-
-
-
cytidine 5'-monophosphosialic acid synthetase
-
-
-
-
cytidine 5-monophosphate N-acetylneuraminic acid synthetase
-
-
-
-
cytidine 5-monophosphate N-acetylneuraminic acid synthetase
-
-
cytidine monophosphate sialic acid synthetase
-
-
cytidine monophosphate-N-acetylneuraminic acid synthetase
-
-
-
-
cytidine monophosphate-sialic acid synthetase
Q99KK2
-
cytidine monophospho-sialic acid synthetase
-
-
-
-
cytidine monophosphoacetylneuraminic synthetase
-
-
-
-
cytidine monophosphosialate pyrophosphorylase
-
-
-
-
cytidine monophosphosialate synthetase
-
-
-
-
cytidyltransferase, acylneuraminate
-
-
-
-
cytidylyltransferase, acetylneuraminate
-
-
-
-
hCSS
-
-
mCSS
-
-
N-Acetyl-neuraminic acid cytidylyltransferase
-
-
sialate cytidylyltransferase
-
-
-
-
sialic acid cytidylyltransferase
-
-
CAS REGISTRY NUMBER
COMMENTARY
9067-82-7
-
ORGANISM
COMMENTARY
LITERATURE
SEQUENCE CODE
SEQUENCE DB
SOURCE
ATCC27405; strain ATCC 27405
-
-
Manually annotated by BRENDA team
Clostridium thermocellum ATCC27405
ATCC27405
-
-
Manually annotated by BRENDA team
wild-type and glycosylation mutant LEC29.Lec32, the mutation resides in either the structural gene encoding CMP-NeuAc synthetase or in a gene that regulates the prpduction of the active enzyme
-
-
Manually annotated by BRENDA team
isoform Cmas1
UniProt
Manually annotated by BRENDA team
isoform Cmas2
UniProt
Manually annotated by BRENDA team
strain K-235
-
-
Manually annotated by BRENDA team
strain K1
-
-
Manually annotated by BRENDA team
Escherichia coli K-235
K-235
-
-
Manually annotated by BRENDA team
Escherichia coli K-235
strain K-235
-
-
Manually annotated by BRENDA team
strain K1
-
-
Manually annotated by BRENDA team
Escherichia coli O18:K1
O18:K1
-
-
Manually annotated by BRENDA team
strain 35000
-
-
Manually annotated by BRENDA team
Haemophilus ducreyi 35000
strain 35000
-
-
Manually annotated by BRENDA team
Mannheimia haemolytica A2
A2
-
-
Manually annotated by BRENDA team
Mannheimia haemolytica A2
strain A2
-
-
Manually annotated by BRENDA team
recombinant
-
-
Manually annotated by BRENDA team
serogroup B
-
-
Manually annotated by BRENDA team
Neisseria meningitidis 406Y
406Y
Uniprot
Manually annotated by BRENDA team
recombinant
-
-
Manually annotated by BRENDA team
Pasteurella multocida ATCC 15742
-
-
-
Manually annotated by BRENDA team
Photobacterium leiognathi JT-SHIZ-145
-
-
-
Manually annotated by BRENDA team
serotype V strain2603 V/R
Uniprot
Manually annotated by BRENDA team
group B, high-producing type Ib strain
-
-
Manually annotated by BRENDA team
Streptococcus sp. Ib
group B, high-producing type Ib strain
-
-
Manually annotated by BRENDA team
hog
-
-
Manually annotated by BRENDA team
GENERAL INFORMATION
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
physiological function
-
CMP-sialic acid synthetase is a key enzyme in the biosynthesis of the capsular polysaccharides required for bacterial infection
SUBSTRATE
PRODUCT                      
REACTION DIAGRAM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
(Substrate)
LITERATURE
(Substrate)
COMMENTARY
(Product)
LITERATURE
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
4-O-methyl-N-acetylneuraminate + CTP
CMP-4-O-methyl-N-acetylneuraminate + diphosphate
show the reaction diagram
-
-
-
?
4-O-methyl-N-acetylneuraminate + CTP
CMP-4-O-methyl-N-acetylneuraminate + diphosphate
show the reaction diagram
-
no activity
-
-
-
CDP + N-acetylneuraminate
phosphate + CMP-N-acetylneuraminate
show the reaction diagram
-
-
-
-
-
CDP + N-acetylneuraminate
phosphate + CMP-N-acetylneuraminate
show the reaction diagram
-
-
-
-
?
CDP + N-acetylneuraminate
phosphate + CMP-N-acetylneuraminate
show the reaction diagram
-
no activity with CDP
-
-
-
CDP + N-acetylneuraminate
phosphate + CMP-N-acetylneuraminate
show the reaction diagram
Mannheimia haemolytica A2
-
-
-
-
?
CDP + N-acetylneuraminate
phosphate + CMP-N-acetylneuraminate
show the reaction diagram
Escherichia coli O18:K1
-
-
-
-
-
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
show the reaction diagram
-
-
-
-
?
CTP + (N-4-O-)diacetylneuraminic acid
diphosphate + CMP-N-4-O-diacetylneuraminate
show the reaction diagram
-
(N-4-O-)diacetylneuraminic acid is 46% effective compared to N-acylneuraminate
-
-
?
CTP + 2-keto-3-deoxy-D-glycero-D-galacto-nononic acid
diphosphate + CMP-2-keto-3-deoxy-D-glycero-D-galacto-nononic acid
show the reaction diagram
Q90WG6
enzyme shows high activity towards 2-keto-3-deoxy-D-glycero-D-galacto-nononic acid and towards N-acetylneuraminate
-
-
?
CTP + 2-keto-3-deoxy-D-glycero-D-galacto-nononic acid
diphosphate + CMP-2-keto-3-deoxy-D-glycero-D-galacto-nononic acid
show the reaction diagram
-, Q99KK2
recombinant enzyme shows 15times lower activity towards 2-keto-3-deoxy-D-glycero-D-galacto-nononic acid than towards N-acetylneuraminate
-
-
?
CTP + 3-deoxy-D-galacto-2-octulosonic acid
diphosphate + CMP-3-deoxy-D-galacto-2-octulosonic acid
show the reaction diagram
-
-
-
-
?
CTP + 5-deaminoneuraminate
diphosphate + CMP-5-deaminoneuraminate
show the reaction diagram
-, H9BFW7, Q0E671
-
-
-
?
CTP + 8-O-methyl-N-acetylneuraminate
diphosphate + CMP-(8-O-methyl)-N-acetylneuraminate
show the reaction diagram
-
-
-
-
?
CTP + deaminoneuraminic acid
?
show the reaction diagram
-
-
-
-
?
CTP + N-(N-benzyloxycarbonyl-glycyl)-muramic acid
diphosphate + CMP-N-(N-benzyloxycarbonyl-glycyl)-muramic acid
show the reaction diagram
-
-
-
-
?
CTP + N-acetyl-neuraminate
diphosphate + CMP-N-acetylneuraminate
show the reaction diagram
-, H9BFW7, Q0E671
-
-
-
?
CTP + N-acetyl-neuraminic acid
CMP-Neu5Ac + diphosphate
show the reaction diagram
-
-
-
-
?
CTP + N-acetylmuramic acid
diphosphate + CMP-N-acetylmuramic acid
show the reaction diagram
-
-
-
-
?
CTP + N-acetylneuraminate
diphosphate + CMP-N-acetylneuraminate
show the reaction diagram
-
-
-
-
?
CTP + N-acetylneuraminate
diphosphate + CMP-N-acetylneuraminate
show the reaction diagram
-
-
-
-
?
CTP + N-acetylneuraminate
diphosphate + CMP-N-acetylneuraminate
show the reaction diagram
-
-
-
-
?
CTP + N-acetylneuraminate
diphosphate + CMP-N-acetylneuraminate
show the reaction diagram
Photobacterium leiognathi JT-SHIZ-145
-
-
-
-
?
CTP + N-acetylneuraminate
diphosphate + CMP-N-acetylneuraminate
show the reaction diagram
Pasteurella multocida ATCC 15742
-
-
-
-
?
CTP + N-acylneuraminate
diphosphate + CMP-N-acylneuraminate
show the reaction diagram
-
-
-
-
-
CTP + N-acylneuraminate
diphosphate + CMP-N-acylneuraminate
show the reaction diagram
-, Q99KK2
-
-
-
-
CTP + N-acylneuraminate
diphosphate + CMP-N-acylneuraminate
show the reaction diagram
-
-
-
-
-
CTP + N-acylneuraminate
diphosphate + CMP-N-acylneuraminate
show the reaction diagram
-
-
-
-
?
CTP + N-acylneuraminate
diphosphate + CMP-N-acylneuraminate
show the reaction diagram
-
-
-
-
-
CTP + N-acylneuraminate
diphosphate + CMP-N-acylneuraminate
show the reaction diagram
-
-
-
-
?
CTP + N-acylneuraminate
diphosphate + CMP-N-acylneuraminate
show the reaction diagram
-
-
-
-
?
CTP + N-acylneuraminate
diphosphate + CMP-N-acylneuraminate
show the reaction diagram
-
-
-
-
-
CTP + N-acylneuraminate
diphosphate + CMP-N-acylneuraminate
show the reaction diagram
-
-
-
-
?
CTP + N-acylneuraminate
diphosphate + CMP-N-acylneuraminate
show the reaction diagram
-
-
-
-
-
CTP + N-acylneuraminate
diphosphate + CMP-N-acylneuraminate
show the reaction diagram
-
-
-
r
CTP + N-acylneuraminate
diphosphate + CMP-N-acylneuraminate
show the reaction diagram
-
-
-
-
r
CTP + N-acylneuraminate
diphosphate + CMP-N-acylneuraminate
show the reaction diagram
-
-
-
-
-
CTP + N-acylneuraminate
diphosphate + CMP-N-acylneuraminate
show the reaction diagram
-
-
-
-
?
CTP + N-acylneuraminate
diphosphate + CMP-N-acylneuraminate
show the reaction diagram
-
-
-
-
?
CTP + N-acylneuraminate
diphosphate + CMP-N-acylneuraminate
show the reaction diagram
P0A0Z8
-
-
-
-
CTP + N-acylneuraminate
diphosphate + CMP-N-acylneuraminate
show the reaction diagram
-
-
-
-
-
CTP + N-acylneuraminate
diphosphate + CMP-N-acylneuraminate
show the reaction diagram
-
-
-
-
?
CTP + N-acylneuraminate
diphosphate + CMP-N-acylneuraminate
show the reaction diagram
P0A0Z8
-
-
-
?
CTP + N-acylneuraminate
diphosphate + CMP-N-acylneuraminate
show the reaction diagram
-
-
-
-
?
CTP + N-acylneuraminate
diphosphate + CMP-N-acylneuraminate
show the reaction diagram
-
-
-
-
ir
CTP + N-acylneuraminate
diphosphate + CMP-N-acylneuraminate
show the reaction diagram
-
-
-
-
?
CTP + N-acylneuraminate
diphosphate + CMP-N-acylneuraminate
show the reaction diagram
-
-
-
-
?
CTP + N-acylneuraminate
diphosphate + CMP-N-acylneuraminate
show the reaction diagram
Q90WG6
-
-
-
-
CTP + N-acylneuraminate
diphosphate + CMP-N-acylneuraminate
show the reaction diagram
-
-
-
-
?
CTP + N-acylneuraminate
diphosphate + CMP-N-acylneuraminate
show the reaction diagram
-
-
-
-
-
CTP + N-acylneuraminate
diphosphate + CMP-N-acylneuraminate
show the reaction diagram
-
-
-
-
?
CTP + N-acylneuraminate
diphosphate + CMP-N-acylneuraminate
show the reaction diagram
-
-
-
-
-
CTP + N-acylneuraminate
diphosphate + CMP-N-acylneuraminate
show the reaction diagram
-
-
-
-
?
CTP + N-acylneuraminate
diphosphate + CMP-N-acylneuraminate
show the reaction diagram
-
-
-
-
?
CTP + N-acylneuraminate
diphosphate + CMP-N-acylneuraminate
show the reaction diagram
-
-
-
-
?
CTP + N-acylneuraminate
diphosphate + CMP-N-acylneuraminate
show the reaction diagram
-
-
-
-
?
CTP + N-acylneuraminate
diphosphate + CMP-N-acylneuraminate
show the reaction diagram
-
-
-
-
-
CTP + N-acylneuraminate
diphosphate + CMP-N-acylneuraminate
show the reaction diagram
-
-
-
-
?
CTP + N-acylneuraminate
diphosphate + CMP-N-acylneuraminate
show the reaction diagram
-
-
-
-
r
CTP + N-acylneuraminate
diphosphate + CMP-N-acylneuraminate
show the reaction diagram
-
-
-
-
?
CTP + N-acylneuraminate
diphosphate + CMP-N-acylneuraminate
show the reaction diagram
-
reaction is theoretically reversible, even though the ratio between the velocities in the anabolic and catabolic direction is 1000:1
-
-
r
CTP + N-acylneuraminate
diphosphate + CMP-N-acylneuraminate
show the reaction diagram
-
reaction is freely reversible and the ratio of the velocities in synthesis and degradation direction is 3:1
-
-
r
CTP + N-acylneuraminate
diphosphate + CMP-N-acylneuraminate
show the reaction diagram
-
production of CMP-N-acetylneuraminate is required for the synthesis of sialylated glycoconjugates
-
-
?
CTP + N-acylneuraminate
diphosphate + CMP-N-acylneuraminate
show the reaction diagram
-
the enzyme produces CMP-N-acetylneuraminate, the nucleotide sugar donor used by sialyltransferase
-
-
?
CTP + N-acylneuraminate
diphosphate + CMP-N-acylneuraminate
show the reaction diagram
-
the enzyme is involved in synthesis of capsular polysaccharide, the recombinant enzyme can function in K1 capsular polysaccharide biosynthesis in Escherichia coli
-
-
?
CTP + N-acylneuraminate
diphosphate + CMP-N-acylneuraminate
show the reaction diagram
-
the lec32 mutation reduces CMP-NeuAc synthetase activity to undetectable levels and reduces Neu-Ac on glycoproteins and glycolipids by 95%
-
-
?
CTP + N-acylneuraminate
diphosphate + CMP-N-acylneuraminate
show the reaction diagram
P0A0Z8
the enzyme is involved in the production of activated sialic acids
-
-
?
CTP + N-acylneuraminate
diphosphate + CMP-N-acylneuraminate
show the reaction diagram
-
neuroinvasive and septicaemia-causing pathogens often display a polysialic acid capsule that is involved in invasive behaviour. N-Acetylneuraminate is the basic monomer of polysialic acid. The activated form CMP-N-acylneuraminate is synthesized by N-acylneuraminate cytidylyltransferase
-
-
?
CTP + N-acylneuraminate
diphosphate + CMP-N-acylneuraminate
show the reaction diagram
-
the enzyme is essential in capsular polysaccharide biosynthesis, activates N-acetylneuraminate for transfer to the nascent capsular polysaccharide
-
-
?
CTP + N-acylneuraminate
diphosphate + CMP-N-acylneuraminate
show the reaction diagram
-
the product of the reaction CMP-N-acylneuraminate is the substrate for the sialylation of glycoconjugates by sialyltransferases
-
-
?
CTP + N-acylneuraminate
diphosphate + CMP-N-acylneuraminate
show the reaction diagram
-
changes in the level of sialylation during development are intimately related to variations in the expression of the enzyme, at least in brain, heart, kidney, stomach, intestine and lung
-
-
?
CTP + N-acylneuraminate
diphosphate + CMP-N-acylneuraminate
show the reaction diagram
-
CMP-NeuAc is essential for the formation of capsule polysialylate for strain K1
-
-
?
CTP + N-acylneuraminate
diphosphate + CMP-N-acylneuraminate
show the reaction diagram
-
enzyme is equally active with either of the three substrates: N-acylneuraminate, N-glycolylneuraminate and deaminoneuraminic acid
-
-
?
CTP + N-acylneuraminate
diphosphate + CMP-N-acylneuraminate
show the reaction diagram
-
enzyme shows pronounced specificity for N-acylneuraminate
-
-
?
CTP + N-acylneuraminate
diphosphate + CMP-N-acylneuraminate
show the reaction diagram
Streptococcus sp. Ib
-
-, the enzyme is essential in capsular polysaccharide biosynthesis, activates N-acetylneuraminate for transfer to the nascent capsular polysaccharide
-
-
?
CTP + N-acylneuraminate
diphosphate + CMP-N-acylneuraminate
show the reaction diagram
Escherichia coli K-235
-
-
-
-
?
CTP + N-acylneuraminate
diphosphate + CMP-N-acylneuraminate
show the reaction diagram
Escherichia coli K-235
-
-
-
-
-
CTP + N-acylneuraminate
diphosphate + CMP-N-acylneuraminate
show the reaction diagram
Clostridium thermocellum ATCC27405
-
-
-
-
?
CTP + N-acylneuraminate
diphosphate + CMP-N-acylneuraminate
show the reaction diagram
Haemophilus ducreyi 35000
-
-
-
-
-
CTP + N-acylneuraminate
diphosphate + CMP-N-acylneuraminate
show the reaction diagram
Haemophilus ducreyi 35000
-
the enzyme produces CMP-N-acetylneuraminate, the nucleotide sugar donor used by sialyltransferase
-
-
?
CTP + N-acylneuraminate
diphosphate + CMP-N-acylneuraminate
show the reaction diagram
-
-, CMP-NeuAc is essential for the formation of capsule polysialylate for strain K1
-
-
?
CTP + N-acylneuraminate
diphosphate + CMP-N-acylneuraminate
show the reaction diagram
Mannheimia haemolytica A2
-
-
-
-
-
CTP + N-acylneuraminate
diphosphate + CMP-N-acylneuraminate
show the reaction diagram
Mannheimia haemolytica A2
-
neuroinvasive and septicaemia-causing pathogens often display a polysialic acid capsule that is involved in invasive behaviour. N-Acetylneuraminate is the basic monomer of polysialic acid. The activated form CMP-N-acylneuraminate is synthesized by N-acylneuraminate cytidylyltransferase
-
-
?
CTP + N-acylneuraminate
diphosphate + CMP-N-acylneuraminate
show the reaction diagram
Mannheimia haemolytica A2
-
-
-
-
?
CTP + N-acylneuraminate
diphosphate + CMP-N-acylneuraminate
show the reaction diagram
Escherichia coli O18:K1
-
-
-
-
?
CTP + N-acylneuraminate methyl ester
diphosphate + CMP-N-acylneuraminate methyl ester
show the reaction diagram
Mannheimia haemolytica, Mannheimia haemolytica A2
-
-
-
-
?
CTP + N-azidoacetylmuramic acid
diphosphate + CMP-N-azidoacetylmuramic acid
show the reaction diagram
-
-
-
-
?
CTP + N-azidomuramic acid
diphosphate + CMP-N-azidomuramic acid
show the reaction diagram
-
-
-
-
?
CTP + N-glycolylneuraminate
diphosphate + CMP-N-glycolylneuraminate
show the reaction diagram
-
-
-
-
?
CTP + N-glycolylneuraminate
diphosphate + CMP-N-glycolylneuraminate
show the reaction diagram
-
-
-
r
CTP + N-glycolylneuraminate
diphosphate + CMP-N-glycolylneuraminate
show the reaction diagram
-
-
-
-
-
CTP + N-glycolylneuraminate
diphosphate + CMP-N-glycolylneuraminate
show the reaction diagram
-
-
-
-
?
CTP + N-glycolylneuraminate
diphosphate + CMP-N-glycolylneuraminate
show the reaction diagram
-
-
-
-
?
CTP + N-glycolylneuraminate
diphosphate + CMP-N-glycolylneuraminate
show the reaction diagram
-
-
-
-
-
CTP + N-glycolylneuraminate
diphosphate + CMP-N-glycolylneuraminate
show the reaction diagram
-
-
-
-
?
CTP + N-glycolylneuraminate
diphosphate + CMP-N-glycolylneuraminate
show the reaction diagram
-
-
-
-
-
CTP + N-glycolylneuraminate
diphosphate + CMP-N-glycolylneuraminate
show the reaction diagram
-
-
-
-
?
CTP + N-glycolylneuraminate
diphosphate + CMP-N-glycolylneuraminate
show the reaction diagram
-
-
-
-
?
CTP + N-glycolylneuraminate
diphosphate + CMP-N-glycolylneuraminate
show the reaction diagram
-
-
-
-
-
CTP + N-glycolylneuraminate
diphosphate + CMP-N-glycolylneuraminate
show the reaction diagram
-
-
-
-
?
CTP + N-glycolylneuraminate
diphosphate + CMP-N-glycolylneuraminate
show the reaction diagram
-
-
-
-
?
CTP + N-glycolylneuraminate
diphosphate + CMP-N-glycolylneuraminate
show the reaction diagram
-, H9BFW7, Q0E671
-
-
-
?
CTP + N-glycolylneuraminate
diphosphate + CMP-N-glycolylneuraminate
show the reaction diagram
Photobacterium leiognathi JT-SHIZ-145
-
-
-
-
?
CTP + N-glycolylneuraminate
diphosphate + CMP-N-glycolylneuraminate
show the reaction diagram
Haemophilus ducreyi 35000
-
-
-
-
?
CTP + N-glycolylneuraminic acid
CMP-N-glycolylneuraminate + diphosphate
show the reaction diagram
-
-
-
-
?
CTP + N-glycolylneuraminic acid
CMP-N-glycolylneuraminate + diphosphate
show the reaction diagram
-
-
-
-
?
CTP + N-glycolylneuraminic acid
CMP-N-glycolylneuraminate + diphosphate
show the reaction diagram
-
-
-
-
?
CTP + N-glycolylneuraminic acid
CMP-N-glycolylneuraminate + diphosphate
show the reaction diagram
Clostridium thermocellum ATCC27405
-
-
-
-
?
CTP + N-glycolylneuraminic acid
CMP-N-glycolylneuraminate + diphosphate
show the reaction diagram
Mannheimia haemolytica A2
-
-
-
-
?
CTP + N-hydroxyacetylmuramic acid
diphosphate + CMP-N-hydroxyacetylmuramic acid
show the reaction diagram
-
-
-
-
?
CTP + N-hydroxymuramic acid
diphosphate + CMP-N-hydroxymuramic acid
show the reaction diagram
-
-
-
-
?
CTP + N-methylglycolylneuraminate
diphosphate + CMP-N-methylglycolylneuraminate
show the reaction diagram
-
-
-
-
?
CTP + sialic acid
CMP-sialic acid + diphosphate
show the reaction diagram
-
-
-
-
?
CTP + sialic acid
CMP-sialic acid + diphosphate
show the reaction diagram
-
-
-
-
?
deaminoneuraminic acid + CTP
CMP-KDN + diphosphate
show the reaction diagram
Q9AFG9, -
N-glycolylneuraminic acid (Neu5Gc) and deaminoneuraminic acid (KDN) are also substrates
-
-
?
N-acetyl-4-O-acetylneuraminate + CTP
diphosphate + CMP-N-acetyl-4-O-acetylneuraminate
show the reaction diagram
-
-
-
-
?
N-acetyl-7(8)-O-acetylneuraminate + CTP
diphosphate + CMP-N-acetyl-7(8)-O-acetylneuraminate
show the reaction diagram
-
-
-
-
?
N-acetylneuraminic acid + CTP
CMP-Neu5Ac + diphosphate
show the reaction diagram
-
DmCSAS is shown to be functional both in vitro and in vivo
-
-
?
N-acetylneuraminic acid + CTP
CMP-Neu5Ac + diphosphate
show the reaction diagram
Q9AFG9, -
N-glycolylneuraminic acid (Neu5Gc) and deaminoneuraminic acid (KDN) are also substrates
-
-
?
N-acetylneuraminic acid + CTP
CMP-NeuAc + diphosphate
show the reaction diagram
-
-
-
-
-
N-acetylneuraminic acid + CTP
CMP-NeuAc + diphosphate
show the reaction diagram
-
-
-
-
?
N-acetylneuraminic acid + CTP
CMP-NeuAc + diphosphate
show the reaction diagram
Clostridium thermocellum, Clostridium thermocellum ATCC27405
-
bi-bi catalytic mechanism, enzyme also accepts N-glycolylneuraminic acid (NeuGc) as a substrate
-
-
?
N-chloroacetylneuraminate + CTP
diphosphate + CMP-N-chloroacetylneuraminate
show the reaction diagram
-
-
-
-
?
N-glycolylneuraminic acid + CTP
CMP-N-glycolylneuraminate + diphosphate
show the reaction diagram
Clostridium thermocellum, Clostridium thermocellum ATCC27405
-
-
-
-
-
N-glycolylneuraminic acid + CTP
CMP-Neu5Gc + diphosphate
show the reaction diagram
Q9AFG9, -
N-glycolylneuraminic acid (Neu5Gc) and deaminoneuraminic acid (KDN) are also substrates
-
-
?
TTP + N-acylneuraminate
diphosphate + TMP-N-acylneuraminate
show the reaction diagram
-
-
-
-
?
TTP + N-acylneuraminate
diphosphate + TMP-N-acylneuraminate
show the reaction diagram
-
no activity with TTP
-
-
-
TTP + N-acylneuraminate
diphosphate + TMP-N-acylneuraminate
show the reaction diagram
Mannheimia haemolytica A2
-
-
-
-
?
TTP + N-glycolylneuraminic acid
TMP-N-glycolylneuraminate + diphosphate
show the reaction diagram
Mannheimia haemolytica, Mannheimia haemolytica A2
-
-
-
-
?
UDP + N-acetylneuraminate
phosphate + CMP-N-acetylneuraminate
show the reaction diagram
-
-
-
-
?
UDP + N-acylneuraminate
phosphate + UMP-N-acetylneuraminate
show the reaction diagram
Mannheimia haemolytica, Mannheimia haemolytica A2
-
-
-
-
?
UTP + N-acylneuraminate
diphosphate + UMP-N-acylneuraminate
show the reaction diagram
-
-
-
-
-
UTP + N-acylneuraminate
diphosphate + UMP-N-acylneuraminate
show the reaction diagram
-
-
-
-
?
UTP + N-acylneuraminate
diphosphate + UMP-N-acylneuraminate
show the reaction diagram
-
-
-
-
?
UTP + N-acylneuraminate
diphosphate + UMP-N-acylneuraminate
show the reaction diagram
-
no activity with UTP
-
-
-
UTP + N-acylneuraminate
diphosphate + UMP-N-acylneuraminate
show the reaction diagram
Mannheimia haemolytica A2
-
-
-
-
?
UTP + N-glycolylneuraminic acid
UMP-N-glycolylneuraminate + diphosphate
show the reaction diagram
-
-
-
-
?
fluoroacetylneuraminate + CTP
diphosphate + CMP-N-fluoroacetylneuraminate
show the reaction diagram
-
-
-
-
?
additional information
?
-
-
no activity with N-acetyl-9-O-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
?
-
Q9AFG9, -
SaV CSS is a bifunctional enzyme having both CMP-sialic acid synthetase and acetylhydrolase (acylesterase) activities
-
-
-
additional information
?
-
-
does not accept N-glycolylneuraminic acid and triacetylneuraminic acid as substrates
-
-
-
additional information
?
-
-
does not utilize N-glycolylneuraminic acid as a substrate
-
-
-
additional information
?
-
-, H9BFW7, Q0E671
enzyme is strictly dependent on CTP
-
-
-
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 substrates: 6'-thioneuraminate, 6-ketodeoxyoctulosonate
-
-
-
additional information
?
-
Escherichia coli K-235
-
does not utilize N-glycolylneuraminic acid as a substrate
-
-
-
additional information
?
-
Photobacterium leiognathi JT-SHIZ-145
-
no substrates: 6'-thioneuraminate, 6-ketodeoxyoctulosonate
-
-
-
NATURAL SUBSTRATES
NATURAL PRODUCTS
REACTION DIAGRAM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
(Substrate)
LITERATURE
(Substrate)
COMMENTARY
(Product)
LITERATURE
(Product)
REVERSIBILITY
r=reversible
ir=irreversible
?=not specified
CTP + N-acylneuraminate
diphosphate + CMP-N-acylneuraminate
show the reaction diagram
-
-
-
-
?
CTP + N-acylneuraminate
diphosphate + CMP-N-acylneuraminate
show the reaction diagram
-
production of CMP-N-acetylneuraminate is required for the synthesis of sialylated glycoconjugates
-
-
?
CTP + N-acylneuraminate
diphosphate + CMP-N-acylneuraminate
show the reaction diagram
-
the enzyme produces CMP-N-acetylneuraminate, the nucleotide sugar donor used by sialyltransferase
-
-
?
CTP + N-acylneuraminate
diphosphate + CMP-N-acylneuraminate
show the reaction diagram
-
the enzyme is involved in synthesis of capsular polysaccharide, the recombinant enzyme can function in K1 capsular polysaccharide biosynthesis in Escherichia coli
-
-
?
CTP + N-acylneuraminate
diphosphate + CMP-N-acylneuraminate
show the reaction diagram
-
the lec32 mutation reduces CMP-NeuAc synthetase activity to undetectable levels and reduces Neu-Ac on glycoproteins and glycolipids by 95%
-
-
?
CTP + N-acylneuraminate
diphosphate + CMP-N-acylneuraminate
show the reaction diagram
P0A0Z8
the enzyme is involved in the production of activated sialic acids
-
-
?
CTP + N-acylneuraminate
diphosphate + CMP-N-acylneuraminate
show the reaction diagram
-
neuroinvasive and septicaemia-causing pathogens often display a polysialic acid capsule that is involved in invasive behaviour. N-Acetylneuraminate is the basic monomer of polysialic acid. The activated form CMP-N-acylneuraminate is synthesized by N-acylneuraminate cytidylyltransferase
-
-
?
CTP + N-acylneuraminate
diphosphate + CMP-N-acylneuraminate
show the reaction diagram
-
the enzyme is essential in capsular polysaccharide biosynthesis, activates N-acetylneuraminate for transfer to the nascent capsular polysaccharide
-
-
?
CTP + N-acylneuraminate
diphosphate + CMP-N-acylneuraminate
show the reaction diagram
-
the product of the reaction CMP-N-acylneuraminate is the substrate for the sialylation of glycoconjugates by sialyltransferases
-
-
?
CTP + N-acylneuraminate
diphosphate + CMP-N-acylneuraminate
show the reaction diagram
-
changes in the level of sialylation during development are intimately related to variations in the expression of the enzyme, at least in brain, heart, kidney, stomach, intestine and lung
-
-
?
CTP + N-acylneuraminate
diphosphate + CMP-N-acylneuraminate
show the reaction diagram
-
CMP-NeuAc is essential for the formation of capsule polysialylate for strain K1
-
-
?
CTP + N-acylneuraminate
diphosphate + CMP-N-acylneuraminate
show the reaction diagram
Streptococcus sp. Ib
-
the enzyme is essential in capsular polysaccharide biosynthesis, activates N-acetylneuraminate for transfer to the nascent capsular polysaccharide
-
-
?
CTP + N-acylneuraminate
diphosphate + CMP-N-acylneuraminate
show the reaction diagram
Escherichia coli K-235
-
-
-
-
?
CTP + N-acylneuraminate
diphosphate + CMP-N-acylneuraminate
show the reaction diagram
Haemophilus ducreyi 35000
-
the enzyme produces CMP-N-acetylneuraminate, the nucleotide sugar donor used by sialyltransferase
-
-
?
CTP + N-acylneuraminate
diphosphate + CMP-N-acylneuraminate
show the reaction diagram
-
CMP-NeuAc is essential for the formation of capsule polysialylate for strain K1
-
-
?
CTP + N-acylneuraminate
diphosphate + CMP-N-acylneuraminate
show the reaction diagram
Mannheimia haemolytica A2
-
neuroinvasive and septicaemia-causing pathogens often display a polysialic acid capsule that is involved in invasive behaviour. N-Acetylneuraminate is the basic monomer of polysialic acid. The activated form CMP-N-acylneuraminate is synthesized by N-acylneuraminate cytidylyltransferase
-
-
?
CTP + N-acylneuraminate
diphosphate + CMP-N-acylneuraminate
show the reaction diagram
Mannheimia haemolytica A2
-
-
-
-
?
CTP + sialic acid
CMP-sialic acid + diphosphate
show the reaction diagram
-
-
-
-
?
CTP + sialic acid
CMP-sialic acid + diphosphate
show the reaction diagram
-
-
-
-
?
METALS and IONS
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
Ca2+
-
divalent cation required, at 20 mM 20% as effective as Mg2+
Ca2+
-
no effect
Ca2+
-
33% of the activation with Mg2+
Ca2+
-
maximal activity in presence of 50 mM Mg2+ or Ca2+
Ca2+
-
divalent metal ion is strictly required, with 20 mM Ca2+ 13% of the activation compared to activation by Mg2+
Ca2+
-
partially substitutes Mg2+ in catalysis
Co2+
-
divalent cation required, at 20 mM 10% as effective as Mg2+
Co2+
-
80% of the activation with Mg2+
Co2+
-
divalent metal ion is strictly required, with 20 mM Mn2+ 27% of the activation compared to activation by Mg2+
Co2+
-
66% as effective as Mn2+; can replace Mn2+, 66% as effective as Mn2+ at 10 mM
Co2+
-
partially substitutes Mg2+ in catalysis
Fe2+
-
divalent cation required, at 20 mM 10% as effective as Mg2+
Fe2+
-
10% of the activation with Mg2+
Mg2+
-
-
Mg2+
-
divalent cation required, maximal stimulation at 20 mM
Mg2+
-
required
Mg2+
-
maximal activity at 20 mM or above; required
Mg2+
-
Mg2+ or Mn2+ required, optimal activity with 20-40 mM Mg2+
Mg2+
-
maximal activity at 10 mM
Mg2+
-
maximal activity in presence of 50 mM Mg2+ or Ca2+
Mg2+
-
40 mM, 30% stimulation
Mg2+
-
divalent metal ion is strictly required, maximal activity at 10 mM
Mg2+
-
required, maximal activity at 20 mM
Mg2+
-
maximal activity at 20 mM
Mg2+
-
20 mM; 57% as effective as Mn2+; can replace Mn2+, 57% as effective as Mn2+ at 10 mM
Mg2+
Q9AFG9, -
required for CMP-sialic acid synthetase activity but not for acetylhydrolase activity
Mg2+
-
assay with 20 mM Mg2+
Mg2+
-
required for activity, 20-40 mM Mg2+ is the optimum concentration
Mg2+
-
absolutely required for activity, 20 mM Mg2+ gives the most activity
Mg2+
-
required for activity, maximum activity at 10 mM Mg2+, decreasing at higher Mg2+ concentrations
Mg2+
-
required for activity
Mg2+
-
required, maximum activity above 5 mM
Mg2+
-
enzyme recruits two Mg2+ ions during the catalytic cycle
Mn2+
-
divalent cation required, at 20 mM 20% as effective as Mg2+
Mn2+
-
Mg2+ or Mn2+ required
Mn2+
-
70% of the activity with Mg2+
Mn2+
-
divalent metal ion is strictly required, with 20 mM Mn2+ 28% of the activation compared to activation by Mg2+
Mn2+
-
20 mM, 40% of the activation with Mg2+
Mn2+
-
10% of the activity with Mg2+
Mn2+
-
absolute requirement for a divalent cation for activity, maximum activity at 10 mM Mn2+; the enzyme has an absolute requirement for a divalent cation for activity and reaches maximum activity in the presence of 10 mM Mn2+
Mn2+
Q9AFG9, -
required for CMP-sialic acid synthetase activity but not for acetylhydrolase activity, maximum activity with 4 mM MnCl
Mn2+
-
required for activity, can be replaced by Mg2+, Co2+, and Zn2+ in decreasing order
Mn2+
-
required for activity, Mn2+ is only 40% as effective as Mg2+ at 20 mM
Mn2+
-
partially substitutes Mg2+ in catalysis
Mn2+
-
required for activity
Zn2+
-
62% as effective as Mn2+; can replace Mn2+, 62% as effective as Mn2+ at 10 mM
Mn2+
-
5% of the activity in presence of Mg2+
additional information
-
Ca2+ shows no effect on enzyme activity
INHIBITORS
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
(NH4)2SO4
-
1.0 M, 83% inhibition, reversible by desalting
2'-deoxy-CTP
-
0.3 mM; 44% inhibition
2-deoxy-2,3-dehydro-N-acetylneuraminate
-
4.6 mM, 33% inhibition
4-(hydroxymercuri) benzoic acid
-
-
4-(hydroxymercuri)benzoic acid
-
-
5,5'-dithiobis(2-nitrobenzoic acid)
-
-
5-Mercuri-CTP
-
-
5-mercury-CTP
-
-
AMP
-
10 mM in presence of 3 mM CTP, 43% inhibition
AMP
-
10 mM, 64% inhibition. 20% inhibition by 1 mM
ATP
-
10 mM, 60% inhibition
Ba2+
-
20 mM, less than 30% inhibition
Ca2+
-
inhibits when incubated in presence of Mg2+ at the same concentration
Ca2+
-
20 mM in presence of 10 mM Mg2+, more than 90% inhibition
Ca2+
-
95% inhibition of enzyme activity at 10 mM
Cd2+
-
inhibits when incubated in presence of Mg2+ at the same concentration
CDP
-
0.3 mM, 45% inhibition
CDP
-
10 mM in presence of 3 mM CTP, 73% inhibition
CDP
-
1 mM, 49% inhibition
CDP
-
1 mM, 37C, pH 8.5, 15 min, 14% inhibition
CMP
-
0.3 mM, 57% inhibition
CMP
-
10 mM in presence of 3 mM CTP, 89% inhibition
CMP
-
noncompetitive
CMP
-
1 mM, 43% inhibition
CMP
-
50 mM, 58% inhibition
CMP-N-acylneuraminate
-
5 mM, 60% inhibition
CMP-N-acylneuraminate
-
-
Co2+
-
inhibits when incubated in presence of Mg2+ at the same concentration
Co2+
-
20 mM in presence of 10 mM Mg2+, more than 70% inhibition
CTP
-
at high concentrations
Cu2+
-
inhibits when incubated in presence of Mg2+ at the same concentration
Cu2+
-
1 mM, 21% inhibition
Cu2+
-
10 mM, 98% inhibition
Cu2+
-
20 mM in presence of 10 mM Mg2+, more than 90% inhibition
Cu2+
-
inhibition in presence of Mg2+
Cu2+
-
97% inhibition of enzyme activity at 10 mM
cytidine
-
60 mM, 37% inhibition
diphosphate
-
1 mM, 46% inhibition
DTT
-
in the presence of 0.01 M DTT enzyme shows 20% activity loss
EDTA
Q9AFG9, -
enzyme is inactive in the presence of 5 mM EDTA
EDTA
-
treatment with EDTA results in loss of catalytic activity which can be restored by the addition of either Mg2+ or Mn2+
EDTA
-
complete inhibition
Fe3+
-
10 mM, 50% inhibition
-
Fe3+
-
20 mM in presence of 10 mM Mg2+, more than 90% inhibition
-
Fe3+
-
91% inhibition of enzyme activity at 10 mM
-
GMP
-
20 mM, 64% inhibition
GTP
-
10 mM in presence of 3 mM CTP, 63% inhibition
GTP
-
10 mM, 39% inhibition
Hg2+
-
1 mM, 33% inhibition
Hg2+
-
10 mM, 62% inhibition
Hg2+
-
20 mM in presence of 10 mM Mg2+, more than 90% inhibition
iodoacetate
-
-
Mn2+
-
inhibits when incubated in presence of Mg2+ at the same concentration
Mn2+
-
20 mM in presence of 10 mM Mg2+, more than 90% inhibition
NaF
-
in the presence of 0.06 M NaF enzyme shows 17% activity loss
NaN3
-
5 mM, 70% inhibition
Ni2+
-
20 mM in presence of 10 mM Mg2+, more than 90% inhibition
p-hydroxymercuribenzoic acid
-
-
periodate-oxidized CTP
-
-
-
PHMB
-
1 mM, complete inhibition
saturated sulfo-CDP
-
1 mM, 37C, pH 8.5, 15 min, 55% inhibition
saturated sulfo-UDP
-
1 mM, 37C, pH 8.5, 15 min, 35% inhibition
saturated sulfo-UDP ester
-
1 mM, 37C, pH 8.5, 15 min, 24% inhibition
Sr2+
-
20 mM, less than 30% inhibition
sulfo-CDP
-
1 mM, 37C, pH 8.5, 15 min, 47% inhibition
sulfo-CTP
-
1 mM, 37C, pH 8.5, 15 min, 39% inhibition
sulfo-UDP
-
1 mM, 37C, pH 8.5, 15 min, 6% inhibition
sulfo-UTP
-
1 mM, 37C, pH 8.5, 15 min, 33% inhibition
TTP
-
10 mM in presence of 3 mM CTP, 71% inhibition
UDP-N-acetylglucosamine
-
10 mM, 26% inhibition
UTP
-
10 mM in presence of 3 mM CTP, 75% inhibition
Zn2+
-
inhibits when incubated in presence of Mg2+ at the same concentration
Zn2+
-
10 mM, 98% of maximal activity
Zn2+
-
inhibits in presence of Mg2+
Mn2+
Q9AFG9, -
both catalytic functions are impaired by high concentrations of Mn2+
additional information
-
enzyme is inhibited by thiol-modifying reagents; enzyme is not inhibited by thiol-containing reagents like GSH, mercaptoethanol and cysteine
-
additional information
-
the addition of monovalent cations (K+, Na+) to the reaction mixtures has no effect on the enzyme activity
-
additional information
Q9AFG9, -
no activity is detected when the enzyme is substituted with 5-35 mM Co2+ or Ca2+
-
additional information
-
dithiothreitol has little effect on enzyme activity
-
ACTIVATING COMPOUND
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
2-mercaptoethanol
-
5 mM, activation to 138% of original activity
2-mercaptoethanol
-
20-30 mM, 3fold stimulation with N-acetylneuraminate as substrate, 1.5fold increase in activity with 4-O-methyl-N-acetylneuraminate as substrate
2-mercaptoethanol
-
1 mM is required for optimal activity of both crude and purified enzyme
2-mercaptoethanol
-
maximal activation, 210%, at 1 mM
ATP
-
1 mM, 20% activation
dithiothreitol
-
activates but enzyme is not dependent on
dithiothreitol
-
5 mM, activates to 193% of the original activity
dithiothreitol
-
Dithiothreitol at a concentration of 0.2 mM stimulates enzyme activity by 158%
DTT
-
maximum activity in the presence of 0.2 mM DTT
additional information
-
mercaptoethanol has no stimulatory effect
-
additional information
Q9AFG9, -
DTT has no effect on enzyme activity
-
additional information
-
no enhancement of activity with dithiothreitol, glutathione, or beta-mercaptoethanol
-
additional information
-
no necessity of including a sulfhydryl reagent, such as dithiothreitol, in the assay reaction
-
KM VALUE [mM]
KM VALUE [mM] Maximum
SUBSTRATE
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
0.56
-
2-keto-3-deoxy-D-glycero-D-galacto-nononic acid
-
pH 9.0, 37C
3
-
2-keto-3-deoxy-D-glycero-D-galacto-nononic acid
-
pH 9.0, 37C
2.5
-
4-O-methyl-N-acetylneuraminate
-
pH 7, 37C
1.5
-
8-O-methyl-N-acetylneuraminate
-
mutant S81R, pH 8.5, 37C
5.3
-
8-O-methyl-N-acetylneuraminate
-
wild-type, pH 8.5, 37C
2.1
-
CDP
-
pH 9.0, 37C, reaction with N-acetylneuraminate
0.00046
-
CTP
-
pH 9.0, 37C
0.0106
-
CTP
-
pH 7.1, 25C
0.017
-
CTP
-
wild-type, pH 7.5, 22C
0.026
-
CTP
-
mutant D211A, pH 7.5, 22C
0.031
-
CTP
-
mutant Q104A, pH 7.5, 22C
0.034
-
CTP
-
CMP-sialic acid synthetase
0.035
-
CTP
-
pH 7.1, 37C
0.035
-
CTP
-
at pH 7.1 and 37C
0.04
-
CTP
-
mutant Y179A, pH 7.5, 22C
0.042
-
CTP
-
N-terminal domain of CMP-sialic acid synthetase
0.043
-
CTP
-
N-terminal domain and C-terminal domain of CMP-sialic acid synthetase
0.048
-
CTP
-
mutant R173A, pH 7.5, 22C
0.052
-
CTP
-
mutant F193A, pH 7.5, 22C
0.15
-
CTP
-
mutant F192A, pH 7.5, 22C
0.16
-
CTP
-
wild-type, pH 8.5, 37C
0.22
-
CTP
-
mutant S81R, pH 8.5, 37C
0.24
-
CTP
-
; at 50C, 10 mM MnCl2, 50 mM Tris buffer (pH 9.5)
0.24
-
CTP
-
wild-type, pH 8.5, 37C
0.27
-
CTP
-
mutant N175A, pH 7.5, 22C
0.31
-
CTP
-
pH 9.0, 37C
0.31
-
CTP
-
at 37C
0.36
-
CTP
-
pH 9.0, 37C, reaction with N-glycolylneuraminate
0.59
-
CTP
-
wild-type, pH 8.5, 37C
0.6
-
CTP
-
pH 9.0, 37C
0.6
-
CTP
-
pH 9.0, 37C
0.96
-
CTP
Q9AFG9, -
-
1.3
-
CTP
-
pH 9.0, 37C
1.5
-
CTP
-
pH 8.0, 37C
1.57
-
CTP
-
pH 9.0, 30C
1.7
-
CTP
-
pH 9.3, 30C
1.77
-
CTP
-
pH 9.0, 37C, reaction with N-acetylneuraminate
2.6
-
CTP
-
pH 9.0, 37C, reaction with N-acetylneuraminate methyl ester
0.62
-
deaminoneuraminic acid
Q9AFG9, -
-
1.2
-
N-acetyl-4-O-acetylneuraminate
-
37C, crude enzyme extract
1.3
-
N-acetyl-4-O-acetylneuraminate
-
37C, crude enzyme extract
1.4
-
N-acetyl-4-O-acetylneuraminate
-
37C, crude enzyme extract
1.1
-
N-acetyl-7(8)-O-acetylneuraminate
-
37C, crude enzyme extract
1.6
-
N-acetyl-7(8)-O-acetylneuraminate
-
37C, crude enzyme extract
0.00036
-
N-acetylneuraminate
-
pH 9.0, 37C
0.066
-
N-acetylneuraminate
-
mutant D211A, pH 7.5, 22C
0.068
-
N-acetylneuraminate
-
wild-type, pH 7.5, 22C
0.076
-
N-acetylneuraminate
-
pH 7.1, 25C
0.1
-
N-acetylneuraminate
-
mutant S81R, pH 8.5, 37C
0.11
-
N-acetylneuraminate
-
mutant Y179A, pH 7.5, 22C
0.14
-
N-acetylneuraminate
-
wild-type, pH 8.5, 37C
0.18
-
N-acetylneuraminate
-
wild-type, pH 8.5, 37C
0.22
-
N-acetylneuraminate
-
wild-type, pH 8.5, 37C
0.26
-
N-acetylneuraminate
-
pH 9.0, 37C
0.26
-
N-acetylneuraminate
-
pH 7.1, 37C
0.29
-
N-acetylneuraminate
-
mutant R173A, pH 7.5, 22C
0.34
-
N-acetylneuraminate
-
pH 9.0, 37C
0.34
-
N-acetylneuraminate
-
pH 8.5, 37C
0.38
-
N-acetylneuraminate
-
mutant F192A, pH 7.5, 22C
0.7
-
N-acetylneuraminate
-
pH 9, 37C
0.78
-
N-acetylneuraminate
-
mutant F193A, pH 7.5, 22C
0.867
-
N-acetylneuraminate
-
pH 9.0, 30C
1
-
N-acetylneuraminate
-
pH 9, 37C
1.2
-
N-acetylneuraminate
-
37C, crude enzyme extract
1.2
-
N-acetylneuraminate
-
-
1.3
-
N-acetylneuraminate
-
pH 8.0, 37C
1.4
-
N-acetylneuraminate
-
37C, crude enzyme extract
1.4
-
N-acetylneuraminate
-
-
1.6
-
N-acetylneuraminate
-
pH 9.0, 37C
1.6
-
N-acetylneuraminate
-
pH 9.0, 37C, reaction with TTP
1.6
-
N-acetylneuraminate
-
mutant N175A, pH 7.5, 22C
1.8
-
N-acetylneuraminate
-
pH 9.0, 37C, reaction with CDP
1.82
-
N-acetylneuraminate
-
pH 9.0, 37C, reaction with CTP
1.9
-
N-acetylneuraminate
-
pH 9.0, 37C, reaction with UDP; pH 9.0, 37C, reaction with UTP
2.1
-
N-acetylneuraminate
-
pH 9.3, 30C
2.2
-
N-acetylneuraminate
-
-
2.6
-
N-acetylneuraminate
-
mutant Q104A, pH 7.5, 22C
2.8
-
N-acetylneuraminate
-
pH 9.0, 37C
4
-
N-acetylneuraminate
-
pH 9.0, 37C
7.6
-
N-acetylneuraminate
-
-
1.3
-
N-acetylneuraminate methyl ester
-
pH 9.0, 37C, reaction with CTP
0.13
-
N-acylneuraminate
-
at 50C, 10 mM MnCl2, 50 mM Tris buffer (pH 9.5)
0.13
-
N-acylneuraminate
-
-
0.26
-
N-acylneuraminate
-
at pH 7.1 and 37C
0.34
-
N-acylneuraminate
-
-
0.8
-
N-acylneuraminate
-
pH 9.0, 37C
1.82
-
N-acylneuraminate
-
-
4
-
N-acylneuraminate
-
at 37C
1.2
-
N-glycolylneuraminate
-
37C, crude enzyme extract
1.3
-
N-glycolylneuraminate
-
37C, crude enzyme extract
1.5
-
N-glycolylneuraminate
-
37C, crude enzyme extract
2.3
-
N-glycolylneuraminate
-
pH 9.0, 37C
2.3
-
N-glycolylneuraminate
-
pH 9.0, 37C
2.6
-
N-glycolylneuraminate
-
pH 8.5, 37C
2.9
-
N-glycolylneuraminate
-
pH 9.3, 30C
3
-
N-glycolylneuraminate
-
pH 9.0, 37C, reaction with CTP
4
-
N-glycolylneuraminate
-
mutant S81R, pH 8.5, 37C
6.2
-
N-glycolylneuraminate
-
wild-type, pH 8.5, 37C
0.16
-
N-glycolylneuraminic acid
-
at 50C, 10 mM MnCl2, 50 mM Tris buffer (pH 9.5)
0.35
-
N-glycolylneuraminic acid
Q9AFG9, -
-
2
-
N-methylglycolylneuraminate
-
wild-type, pH 8.5, 37C
-
7
-
N-methylglycolylneuraminate
-
mutant S81R, pH 8.5, 37C
-
5.6
-
Neu5Ac
Q9AFG9, -
-
0.045
-
sialic acid
-
CMP-sialic acid synthetase, substrate N-acetylneuraminic acid
0.063
-
sialic acid
-
N-terminal domain and C-terminal domain of CMP-sialic acid synthetase, substrate N-acetylneuraminic acid
0.07
-
sialic acid
-
N-terminal domain of CMP-sialic acid synthetase, substrate N-acetylneuraminic acid
8.1
-
TTP
-
pH 9.0, 37C, reaction with N-acetylneuraminate
3
-
UDP
-
pH 9.0, 37C, reaction with N-acetylneuraminate
2.7
-
UTP
-
pH 9.0, 37C, reaction with N-acetylneuraminate
TURNOVER NUMBER [1/s]
TURNOVER NUMBER MAXIMUM[1/s]
SUBSTRATE
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
1.9
-
8-O-methyl-N-acetylneuraminate
-
mutant S81R, pH 8.5, 37C
21
-
8-O-methyl-N-acetylneuraminate
-
wild-type, pH 8.5, 37C
0.001
-
CTP
-
mutant D211A, pH 7.5, 22C
0.05
-
CTP
-
mutant Y179A, pH 7.5, 22C
1.8
-
CTP
-
pH 7.1, 25C
2.2
-
CTP
-
mutant Q104A, pH 7.5, 22C
2.57
-
CTP
-
CMP-sialic acid synthetase
3.33
-
CTP
-
; at 50C, 10 mM MnCl2, 50 mM Tris buffer (pH 9.5)
3.87
-
CTP
-
N-terminal domain and C-terminal domain of CMP-sialic acid synthetase
3.89
-
CTP
-
N-terminal domain of CMP-sialic acid synthetase
4.6
-
CTP
-
wild-type, pH 8.5, 37C
5
-
CTP
-
mutant F193A, pH 7.5, 22C
5.2
-
CTP
-
wild-type, pH 8.5, 37C
7
-
CTP
-
mutant R173A, pH 7.5, 22C
8.8
-
CTP
-
mutant S81R, pH 8.5, 37C
10.2
-
CTP
-
wild-type, pH 7.5, 22C
20
-
CTP
-
mutant F192A, pH 7.5, 22C
21
-
CTP
-
wild-type, pH 8.5, 37C
25
-
CTP
-
mutant N175A, pH 7.5, 22C
0.067
-
deaminoneuraminic acid
Q9AFG9, -
-
4.2
-
N-acetylneuraminate
-
wild-type, pH 8.5, 37C
4.8
-
N-acetylneuraminate
-
wild-type, pH 8.5, 37C
7.4
-
N-acetylneuraminate
-
mutant S81R, pH 8.5, 37C
19
-
N-acetylneuraminate
-
wild-type, pH 8.5, 37C
32
-
N-acetylneuraminate
-
pH 8.5, 37C
2.25
-
N-acylneuraminate
-
at 50C, 10 mM MnCl2, 50 mM Tris buffer (pH 9.5)
3.2
-
N-glycolylneuraminate
-
wild-type, pH 8.5, 37C
23
-
N-glycolylneuraminate
-
mutant S81R, pH 8.5, 37C
36
-
N-glycolylneuraminate
-
pH 8.5, 37C
0.68
-
N-glycolylneuraminic acid
Q9AFG9, -
-
1.66
-
N-glycolylneuraminic acid
-
at 50C, 10 mM MnCl2, 50 mM Tris buffer (pH 9.5)
2.9
-
N-methylglycolylneuraminate
-
wild-type, pH 8.5, 37C
-
12
-
N-methylglycolylneuraminate
-
mutant S81R, pH 8.5, 37C
-
16
-
Neu5Ac
Q9AFG9, -
-
2.29
-
sialic acid
-
CMP-sialic acid synthetase, substrate N-acetylneuraminic acid
3.47
-
sialic acid
-
N-terminal domain and C-terminal domain of CMP-sialic acid synthetase, substrate N-acetylneuraminic acid
3.56
-
sialic acid
-
N-terminal domain of CMP-sialic acid synthetase, substrate N-acetylneuraminic acid
kcat/KM VALUE [1/mMs-1]
kcat/KM VALUE [1/mMs-1] Maximum
SUBSTRATE
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
0.55
-
8-O-methyl-N-acetylneuraminate
-
wild-type, pH 8.5, 37C
274252
1.3
-
8-O-methyl-N-acetylneuraminate
-
mutant S81R, pH 8.5, 37C
274252
0.0013
-
CTP
-
mutant Y179A, pH 7.5, 22C
8829
0.072
-
CTP
-
mutant Q104A, pH 7.5, 22C
8829
0.09
-
CTP
-
mutant N175A, pH 7.5, 22C
8829
0.097
-
CTP
-
mutant F193A, pH 7.5, 22C
8829
0.132
-
CTP
-
mutant F192A, pH 7.5, 22C
8829
0.143
-
CTP
-
mutant R173A, pH 7.5, 22C
8829
0.6
-
CTP
-
wild-type, pH 7.5, 22C
8829
19
-
CTP
-
wild-type, pH 8.5, 37C
8829
33
-
CTP
-
wild-type, pH 8.5, 37C
8829
36
-
CTP
-
wild-type, pH 8.5, 37C
8829
40
-
CTP
-
mutant S81R, pH 8.5, 37C
8829
0.0005
-
N-acetylneuraminate
-
mutant Y179A, pH 7.5, 22C
13652
0.0009
-
N-acetylneuraminate
-
mutant Q104A, pH 7.5, 22C
13652
0.0063
-
N-acetylneuraminate
-
mutant F193A, pH 7.5, 22C
13652
0.052
-
N-acetylneuraminate
-
mutant F192A, pH 7.5, 22C
13652
0.148
-
N-acetylneuraminate
-
wild-type, pH 7.5, 22C
13652
0.155
-
N-acetylneuraminate
-
mutant N175A, pH 7.5, 22C
13652
0.23
-
N-acetylneuraminate
-
mutant R173A, pH 7.5, 22C
13652
27
-
N-acetylneuraminate
-
wild-type, pH 8.5, 37C
13652
30
-
N-acetylneuraminate
-
wild-type, pH 8.5, 37C
13652
74
-
N-acetylneuraminate
-
mutant S81R, pH 8.5, 37C
13652
86
-
N-acetylneuraminate
-
wild-type, pH 8.5, 37C
13652
3.8
-
N-glycolylneuraminate
-
wild-type, pH 8.5, 37C
13913
5.9
-
N-glycolylneuraminate
-
mutant S81R, pH 8.5, 37C
13913
1.6
-
N-methylglycolylneuraminate
-
wild-type, pH 8.5, 37C
0
1.7
-
N-methylglycolylneuraminate
-
mutant S81R, pH 8.5, 37C
0
Ki VALUE [mM]
Ki VALUE [mM] Maximum
INHIBITOR
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
0.5
-
CMP
-
pH 9.0, 37C
additional information
-
additional information
-
-
-
IC50 VALUE [mM]
IC50 VALUE [mM] Maximum
INHIBITOR
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
additional information
-
CDP
-
1 mM, 37C, pH 8.5, 15 min, 14% inhibition
additional information
-
saturated sulfo-CDP
-
1 mM, 37C, pH 8.5, 15 min, 55% inhibition
additional information
-
saturated sulfo-UDP
-
1 mM, 37C, pH 8.5, 15 min, 35% inhibition
additional information
-
saturated sulfo-UDP ester
-
1 mM, 37C, pH 8.5, 15 min, 24% inhibition
additional information
-
sulfo-CDP
-
1 mM, 37C, pH 8.5, 15 min, 47% inhibition
additional information
-
sulfo-CTP
-
1 mM, 37C, pH 8.5, 15 min, 39% inhibition
additional information
-
sulfo-UDP
-
1 mM, 37C, pH 8.5, 15 min, 6% inhibition
additional information
-
sulfo-UTP
-
1 mM, 37C, pH 8.5, 15 min, 33% inhibition
SPECIFIC ACTIVITY [µmol/min/mg]
SPECIFIC ACTIVITY MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
0.0017
-
-
mutant enzyme DELTA1-227
0.0113
-
-
wild-type enzyme
0.045
-
-
-
0.165
-
-
30C, pH 9.0
0.2
-
-, H9BFW7, Q0E671
substrate N-glycolylneuraminate, pH 7.5, temperature not specified in the publication
0.3
-
-, H9BFW7, Q0E671
substrate 5-deaminoneuraminate, pH 7.5, temperature not specified in the publication
0.542
-
-
-
0.83
-
-
at pH 7.6
1.58
3.17
-
at pH 9.0
2.7
-
-, H9BFW7, Q0E671
substrate N-glycolylneuraminate, pH 7.5, temperature not specified in the publication
3.3
-
-, H9BFW7, Q0E671
substrate 5-deaminoneuraminate, pH 7.5, temperature not specified in the publication
4.5
-
-, H9BFW7, Q0E671
substrate N-acetylneuraminate, pH 7.5, temperature not specified in the publication
18.25
-
-
-
additional information
-
-
-
additional information
-
-
simple and efficient method for assaying cytidine monophosphate sialic acid synthetase activity using an enzymatic reduced nicotinamide adenine dinucleotide/oxidized nicotinamide adenine dinucleotide converting system
pH OPTIMUM
pH MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
7
-
-
reaction with Mn2+
7.5
-
-
assay at
8
-
-
at an ionic strength ranging between 50-250 mM, in Tris-HCl buffer, glycine-NaOH as well as in phosphate buffer
8
-
-
activity assay
8.5
9
-
activity in presence of Mg2+
8.5
-
-
inhibition assay
9
-
Q9AFG9, -
optimal CMP-sialic acid synthetase activity
9.5
-
-
-
9.5
-
-
; maximum activity
pH RANGE
pH RANGE MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
7
10
-
pH 7.0: about 90% of maximal activity, pH 10.0: about 70% of maximal activity
7
12.5
-
activity decreases dramatically below pH 7.0 and above pH 12.5
7.5
10.5
Q9AFG9, -
enzyme is active in a wide pH-range
7.5
9.5
-
pH 7.5: about 20% of maximal activity, pH 8.0: about 90% of maximal activity, pH 9.5: about 65% of maximal activity
8
13
-
enzyme is active between pH 8-13; in Tris-HCl buffer
TEMPERATURE OPTIMUM
TEMPERATURE OPTIMUM MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
22
-
-
activity assay
37
-
-
assay at
37
-
Q9AFG9, -
assay at
37
-
-
inhibition assay
40
60
-
immobilized enzyme yields maximal activity at 40C which is maintained up to 60C, whereas the activity of the soluble enzyme decreases sharply above 40C
40
-
-
greatest activity is observed at 40C
50
-
-
; maximum activity
TEMPERATURE RANGE
TEMPERATURE MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
25
42
-
4C: activity is undetectable, 25-37C: maximal activity, 42C: 12% of maximal activity
30
50
-
30C: about 40% of maximal activity, 50C: about 90% of maximal activity
33
42
-
50% of maximal activity at 33C and at 42C
37
60
-
enzyme is active between 37C - 60C; in the presence of 0.2 mM dithiothreitol
pI VALUE
pI VALUE MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
5
-
-
isoelectric focusing
5
-
-
isoelectric focusing
5.6
5.9
-
isoelectric focusing
5.9
-
-
isoelectric focusing
6.5
-
-
chromatofocusing
SOURCE TISSUE
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
SOURCE
-, H9BFW7, Q0E671
during the pharyngula stage, expression is restricted to the brain
Manually annotated by BRENDA team
-, H9BFW7, Q0E671
robust expression in the entire central nervous system, the somites, the notochord, and the developing pronephric duct
Manually annotated by BRENDA team
-, H9BFW7, Q0E671
more or less ubiquitously expressed from the end of gastrula through segmentation
Manually annotated by BRENDA team
-
DmCSAS gene is greatly enriched in adult head, suggesting a possible role in the central nervous system
Manually annotated by BRENDA team
-, H9BFW7, Q0E671
robust expression in the entire central nervous system, the somites, the notochord, and the developing pronephric duct
Manually annotated by BRENDA team
-, H9BFW7, Q0E671
robust expression in the entire central nervous system, the somites, the notochord, and the developing pronephric duct
Manually annotated by BRENDA team
additional information
-
no activity in skeletal muscle. The expression the enzyme is tissue-dependent and does not seem to be embryonically determined. Changes in the level of sialylation during development are intimately related to variations in the expression of the enzyme, at least in brain, heart, kidney, stomach, intestine and lung
Manually annotated by BRENDA team
additional information
-
DmCSAS gene is developmentally regulated, with expression first appearing at 12-24 h of embryogenesis, low expression through larval and pubal stages
Manually annotated by BRENDA team
additional information
-, H9BFW7, Q0E671
not detected in skeletal muscle, liver, or kidney
Manually annotated by BRENDA team
LOCALIZATION
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
GeneOntology No.
LITERATURE
SOURCE
-
; a small, but significant population of the mouse CMP-sialic acid synthetase is also present in cytoplasm
Manually annotated by BRENDA team
-
mostly localized in nucleus; vertebrate CSS are usually localised in the nucleus due to a nuclear localisation signal, however in mouse a small proportion of CSS is also present in cytoplasm due to two nuclear export signals
Manually annotated by BRENDA team
-, H9BFW7, Q0E671
;
Manually annotated by BRENDA team
-
unlike all vertebrate CSAS that localize to the nucleus, the DmCSAS is localized to the Golgi compartment
Manually annotated by BRENDA team
additional information
-
replacement of the N-terminal leader sequence of DmCSAS with the human CSAS N-terminal sequence results in the redirection of the chimeric CSAS protein to the nucleus but with concomitant loss of activity
-
Manually annotated by BRENDA team
MOLECULAR WEIGHT
MOLECULAR WEIGHT MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
18300
-
-
C-terminal domain of CMP-sialic acid synthetase, calculated
24800
-
-
gel filtration
25400
-
-
gel filtration
26400
-
-
N-terminal domain of CMP-sialic acid synthetase, calculated
27500
-
-
predicted from cDNA
43000
-
-
SDS-PAGE
44400
-
-
CMP-sialic acid synthetase, calculated
49000
-
-
SDS-PAGE
55800
-
-
gel filtration
56600
-
-
dimer, N-terminal domain of CMP-sialic acid synthetase, determined by gel filtration
87000
-
-
tetramer, C-terminal domain of CMP-sialic acid synthetase, determined by gel filtration
116000
-
-
gel filtration
135000
-
-
gel filtration
158000
-
-
gel filtration
160000
-
-
gel filtration
162000
-
-
gel filtration
175800
-
-
tetramer, CMP-sialic acid synthetase, determined by gel filtration
193400
-
-, H9BFW7, Q0E671
gel filtration
236100
-
-, H9BFW7, Q0E671
gel filtration
additional information
-
-
47000-74000 Da, gel filtration
SUBUNITS
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
?
-
x * 50000, SDS-PAGE
?
-
x * 24892, calculation from nucleotide sequence; x * 24893, mass spectrometry
?
-
x * 69000, SDS-PAGE
?
-
x * 48058, calculation from nucleotide sequence
?
-
x * 27313, electrospray mass spectrometry; x * 33700, SDS-PAGE
?
Escherichia coli O18:K1
-
x * 50000, SDS-PAGE
-
dimer
-
2 * 58000, SDS-PAGE
dimer
-
2 * 25440, dimer or larger oligomer, electrospray mass spectrometry; 2 * 25600, dimer or larger oligomer, SDS-PAGE
dimer
Haemophilus ducreyi 35000
-
2 * 25440, dimer or larger oligomer, electrospray mass spectrometry; 2 * 25600, dimer or larger oligomer, SDS-PAGE
-
monomer
-
1 * 164000, SDS-PAGE
monomer
-
1 * 60000, SDS-PAGE
monomer
Photobacterium leiognathi JT-SHIZ-145
-
1 * 60000, SDS-PAGE
-
pentamer
-, H9BFW7, Q0E671
or tetramer, 5 * 48200, calculated
tetramer
-
4*190000, gel filtration
tetramer
-
dimer of dimers
tetramer
-, H9BFW7, Q0E671
4 * 49600, calculated; or pentamer, 4 * 48200, calculated
trimer
-
3 * 52000, SDS-PAGE
trimer
-
3 * 43000, SDS-PAGE
trimer
Mannheimia haemolytica A2
-
3 * 43000, SDS-PAGE
-
POSTTRANSLATIONAL MODIFICATION
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
additional information
-
no carbohydrates can be detected
Crystallization/COMMENTARY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
the structure of the C-terminal domain of CMP-sialic acid synthetase is solved to 1.9 A resolution
-
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
-
pH STABILITY
pH STABILITY MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
7
12.5
-
activity is drastically reduced below pH 7.0 and above pH 12.5
7
-
-
above, 4C, stable for 4 weeks
7
-
-
most stable at neutral pH
TEMPERATURE STABILITY
TEMPERATURE STABILITY MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
22
-
-
not stable at room temperature for 12 h
25
-
-
unstable even at
25
-
Q9AFG9, -
enzyme is more stable at lower temperatures, 55% of enzyme activity remains after 24 h incubation at 25C
35
-
-
60 min, completely stable
37
70
-
complete loss of activity above 70C, no loss of activity after 24 h incubation at 37C and 50C
37
-
-
reaction rate is proportional to enzyme concentration and constant for at least 45 min. At high enzyme concentrations the reaction rate decreases 10-15% between 45 and 60 min
37
-
-
60 min, about 10% loss of activity
37
-
Q9AFG9, -
enzyme is not stable at 37C, incubation for 1 h decreases enzyme activity by 50%, incubation for 24 h at 37C completely inactivates enzyme activity
40
45
-
the enzyme loses complete activity after 50 and 10 min when stored at 40C and 45C, respectively
40
-
-
60 min, 20% loss of activity
40
-
-
50 min, complete inactivation without stabilizer, completely stable in presence of 1.0 M NaCl, 0.5 M Na2HPO4 or 25% v/v glycerol
45
-
-
60 min, 65% loss of activity, less than 10% loss of activity in presence of 20% w/v glycerol
45
-
-
10 min, complete deactivation
50
-
-
10 min, 85% loss of activity
55
-
-
20 min, complete loss of activity
70
-
-
the enzyme retains activity at 70C
GENERAL STABILITY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
the enzyme is equally stable with or without the presence of glycerol at different temperatures
-
stabilized against thermal inactivation by some salts, NaCl, Na2SO4 or Na2HPO4, or by glycerol
-
enzyme is stable in lyophilized form
-
stability can be improved by 1 mM 2-mercaptoethanol
-
stable in the frozen state, each cycle of freezing and thawing leads to a 15% decrease in activity
-
purified enzyme is sensitive to repeated freezing
-
the enzyme tolerates flash freezing and lyophilization
-
not stable to freezing and thawing, addition of substrates partially stabilizes the purified enzyme to freezing and thawing
-
not stable to lyophilization
-
STORAGE STABILITY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
-20C, stable for at least 1 year
-
-20C, partially purified enzyme can be stored in 43% glycerol or in 50 mM Tris/HCl or potassium phosphate, pH 7.5, for several months without loss of activity
-
4C, above pH 7.0, stable for several weeks
-
4C, at pH 7.0, several weeks, remains stable
-
-20C or -80C, in 50% (v/v) glycerol, several months, remains stable
-
-20C or -80C, the relatively crude preparation is stable to storage as a 50% glycerol solution for at least several months
-
4C, the relatively crude preparation is stable to storage as a 65% ammonium sulfate suspension or 20% glycerol solution for at least several months
-
4C, 7 months, a magnetic nanoparticle-CSS preparation retains almost 100% activity
-
-20C, stable for at least 1 year
-
-20C, 2 weeks, more than 50% loss of activity
-
-80C, completely stable for 8 months
-
4C, pH 7.6, 83% of the original activity is recovered after 4 weeks
-
4C, stable for 3 weeks
-
Purification/COMMENTARY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
Ni-NTA column chromatography
-
wild-type enzyme and mutant enzymes
-
partial
-
recombinant proteins are expressed in Escherichia coli BL21DE3 cells and purified by affinity chromatography utilizing the StrepII-Tag, further purified on a Superdex 200 HR 10/30 columnn
-
by column chromatography on Sephacryl S-200 (gel filtration), Yellow-86-Agarose (affinity) and Phenyl-Sepharose (hydrophobic affinity)
-
C-His6-tagged CSS is purified using a nickel-nitrilotriacetic acid-agarose (Ni2+-NTA-agarose) affinity column
Q9AFG9, -
Cloned/COMMENTARY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
cloned in Escherichia coli XL-10; expressed in Escherichia coli strain BL21
-
transfection of NIH-3T3 and EPC cells; transfection of NIH-3T3 and EPC cells
-, H9BFW7, Q0E671
DmCSAS is cloned into the baculovirus vector pBlueBac4.5 (pBlueBac-DmCSAS4) which is used to transfect LEC29.Lec32 cells
-
a series of deletions from the 3'-end of the CMP-NeuAc synthetase coding region is constructed and expressed in Escherichia coli
-
expression in Escherichia coli
-
the gene is cloned into a T7 expression vector, the protein is expressed in Escherichia coli
-
human CMP-N-acetylneuraminic acid synthetase is expressed in tobacco BY2 suspension-cultured cells, enzyme is demonstrated to be active
-
cloning is achieved by complementation of the Chineses hamster ovary lec32 mutation that causes a deficiency in CMP-N-acetylneuraminate synthetase activity, it also causes polysialic acid to be expressed in the capsule of the CMP-Neu5Ac synthetase negative Escherichia coli mutant EV5; expression in CHO cells or Escherichia coli
-
expressed in CHO cells, HeLa cells, and NIH-3T3 cells; expression in CHO cells
-
the N-terminal domain, residues 39-267, and the C-terminal domain, residues 267-432, of CMP-sialic acid synthetase, and CMP-sialic acid synthetase, residues 39-432, are cloned into a pET22b-Strep vector
-
expression in Escherichia coli
-
high-level expression in Escherichia coli
-
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
-
expressed in Escherichia coli and CHO cells
-
expression in Escherichia coli
-
expression in Escherichia coli
-
cloned in Escherichia coli as a His6-tagged fusion protein
Q9AFG9, -
expression in Escherichia coli, enzyme can function in K1 capsular polysaccharide biosynthesis in Escherichia coli
-
ENGINEERING
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
DELTA1-227
-
mutant enzyme shows no CMP-N-acetylneuraminic acid synthetase activity, mutant enzyme is able to hydrolyze platelet activating factor
DELTA1-229
-
mutant enzyme shows no CMP-N-acetylneuraminic acid synthetase activity, mutant enzyme is not able to hydrolyze platelet activating factor
DELTA230-418
-
mutant enzyme shows 43% of the wild-type activity with CTP and N-acylneuraminate as substrates, decrease in stability compared to wild-type enzyme
DELTA247-418
-
mutant enzyme shows 15% of the wild-type activity with CTP and N-acylneuraminate as substrates, decrease in stability compared to wild-type enzyme
DELTA340-418
-
mutant enzyme shows 65% of the wild-type activity with CTP and N-acylneuraminate as substrates, as stable as wild-type enzyme
DELTA384-418
-
mutant enzyme shows 31% of the wild-type activity with CTP and N-acylneuraminate as substrates, as stable as wild-type enzyme
DELTA1-227
-
mutant enzyme shows no CMP-N-acetylneuraminic acid synthetase activity, mutant enzyme is able to hydrolyze platelet activating factor
-
DELTA340-418
-
mutant enzyme shows 65% of the wild-type activity with CTP and N-acylneuraminate as substrates, as stable as wild-type enzyme
-
DELTA384-418
-
mutant enzyme shows 31% of the wild-type activity with CTP and N-acylneuraminate as substrates, as stable as wild-type enzyme
-
DELTA408-424
-
deletion mutant devoid of the second putative nuclear export signal, by immunofluorescent microscopy it is shown that this deletion mutant has an increased nuclear localisation combined with a decreased cytoplasmic localisation
DELTA61-69
-
deletion mutant devoid of the first putative nuclear export signal, by immunofluorescent microscopy it is shown that is deletion mutant has an increased nuclear localisation combined with a decreased cytoplasmic localisation
R199A
-
inactive mutant protein
R199A/R202A
-
inactive mutant protein
R201A
-
mutant enzyme is active at moderately reduced level
R202A
-
mutant enzyme shows drastically reduced activity
D209A
-
almost complete loss of activity. Residue involved in metal binding
D211A
-
dramatic loss of activity. Residue involved in metal binding
F192A
-
8fold increase on Km value for CTP
F193A
-
3fold increase on Km value for CTP
K142A
-
10000fold reduction in kcat with an insignificant, fourfold, rise in Km for CTP; dramatic loss of activity. Residue involved in metal binding
N175A
-
16fold increases in the Km value for CTP and 23fold for N-acetylneuraminate
Q104A
-
40fold inccrease in Km value 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
Q163A
-
mutant displays improved substrate promiscuity
R173A
-
38fold increase in Km value for N-acetylneuraminate
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
additional information
-, H9BFW7, Q0E671
presence of four basic clusters BC1-BC4 in the protein sequence. Deletion of BC3 does not impair nuclear import of isoform Cmas1, deletion of BC1 or BC2, individually or in combination, entail retention in the cytoplasm. neither the deletion of BC1 nor BC2 alters enzymatic activity
additional information
-
in contrast to its mammalian counterparts DmCSAS protein begins with an N-terminal sequence rich in hydrophobic amino acids characteristic of a signal/anchoring sequence; multiple alignment among the Drosophila melanogaster, human, mouse, Escherichia coli and Neisseria meningitidis CSAS enzymes shows that all proteins are homologous over the length of the catalytic domain; replacement of the N-terminal leader sequence of DmCSAS with the human CSAS N-terminal sequence results in the redirection of the chimeric CSAS protein to the nucleus but with concomitant loss of activity
DELTA396-418
-
mutant enzyme shows 38% of the wild-type activity with CTP and N-acylneuraminate as substrates, as stable as wild-type enzyme
additional information
-
a series of deletions from the 3'-end of the CMP-NeuAc synthetase coding region is constructed and expressed in Escherichia coli. As a result, the catalytic domain required for CMP-NeuAc synthetase is found to be in the N-terminal half consisting of amino acids 1229. The C-terminal half consisting of amino acids 228418 exhibits platelet-activating factor acetylhydrolase activity
DELTA396-418
-
mutant enzyme shows 38% of the wild-type activity with CTP and N-acylneuraminate as substrates, as stable as wild-type enzyme
-
additional information
-
a series of deletions from the 3'-end of the CMP-NeuAc synthetase coding region is constructed and expressed in Escherichia coli. As a result, the catalytic domain required for CMP-NeuAc synthetase is found to be in the N-terminal half consisting of amino acids 1229. The C-terminal half consisting of amino acids 228418 exhibits platelet-activating factor acetylhydrolase activity
-
K198A
-
mutant enzyme is active at moderately reduced level
additional information
-
by deletion mutant analysis it is demonstrated that mouse CSS contains two nuclear export signals which regulate the transport of the protein from nucleus towards cytosol; the two nuclear export signals of mouse CSS are conserved among mammals and fish CSSs but they are not present in bacteria or insect CSS
Y179A
-
200fold decrease in kcat value
additional information
Q9AFG9, -
amino acid sequence alignment of SaV CSS indicate that the C-terminus belongs to the newly discovered SGNH-hydrolase subfamily of serine esterases/lipases
APPLICATION
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
synthesis
-
the enzyme is used for preparative synthesis of CMP-N-acylneuraminate. Optimization of the culture conditions for the production of the enzyme and the detection of a colony variant of Escherichia coli K-235 that is an even better producer of the synthetase
synthesis
Escherichia coli K-235
-
the enzyme is used for preparative synthesis of CMP-N-acylneuraminate. Optimization of the culture conditions for the production of the enzyme and the detection of a colony variant of Escherichia coli K-235 that is an even better producer of the synthetase
-
agriculture
-
this approach enables plant cells to produce glycoproteins with mammalian-type N-glycans
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
-
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