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Literature summary extracted from

  • Hinderlich, S.; Stäsche, R.; Zeitler, R.; Reutter, W.
    A bifunctional enzyme catalyzes the first two steps in N-acetylneuraminic acid biosynthesis of rat liver. Purification and characterization of UDP-N-acetylglucosamine 2-epimerase/N-acetylmannosamine kinase (1997), J. Biol. Chem., 272, 24313-24318.
    View publication on PubMed

Cloned(Commentary)

EC Number Cloned (Comment) Organism
3.2.1.183
-
Rattus norvegicus

General Stability

EC Number General Stability Organism
2.7.1.60 hexameric enzyme is completely stable with 0.1 mM UDP Rattus norvegicus

Inhibitors

EC Number Inhibitors Comment Organism Structure
3.2.1.183 CMP-N-acetylneuraminic acid
-
Rattus norvegicus
5.1.3.14 CMP-N-acetylneuraminic acid
-
Rattus norvegicus

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
2.7.1.60 0.093
-
N-acetyl-D-mannosamine pH 7.5, 37°C, hexamer Rattus norvegicus
2.7.1.60 0.121
-
N-acetyl-D-mannosamine pH 7.5, 37°C, dimer Rattus norvegicus
2.7.1.60 1.18
-
ATP pH 7.5, 37°C, hexamer Rattus norvegicus
2.7.1.60 1.67
-
ATP pH 7.5, 37°C, dimer Rattus norvegicus
3.2.1.183 11
-
UDP-N-acetyl-alpha-D-glucosamine pH 7.5, 37°C, hexamer Rattus norvegicus
5.1.3.14 0.011
-
UDP-N-acetyl-D-glucosamine bifunctional enzyme EC5.1.3.14/EC2.7.1.60 Rattus norvegicus

Localization

EC Number Localization Comment Organism GeneOntology No. Textmining
2.7.1.60 cytosol
-
Rattus norvegicus 5829
-
3.2.1.183 cytosol
-
Rattus norvegicus 5829
-
5.1.3.14 cytosol
-
Rattus norvegicus 5829
-

Molecular Weight [Da]

EC Number Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
2.7.1.60 75000
-
gel filtration, SDS-PAGE Rattus norvegicus
2.7.1.60 150000
-
gel filtration, ion of the monomeric 75000 Da protein self associate as dimer Rattus norvegicus
2.7.1.60 450000
-
gel filtration, ion of the monomeric 75000 Da protein self associate as hexamer Rattus norvegicus
3.2.1.183 75000
-
monomer, SDS-PAGE, bifunctional enzymatic activity Rattus norvegicus
3.2.1.183 450000
-
gel filtration, dimer. Dimer does not show UDP-GlcNAc 2-epimerase activity. Incubation of the dimer with UDP-N-acetylglucosamine leads to reassembly of the fully active hexamer Rattus norvegicus
3.2.1.183 450000
-
gel filtration, hexamer, bifunctional enzyme complex Rattus norvegicus
5.1.3.14 450000
-
gel filtration Rattus norvegicus

Natural Substrates/ Products (Substrates)

EC Number Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
2.7.1.60 ATP + N-acetyl-D-mannosamine Rattus norvegicus
-
ADP + N-acetyl-D-mannosamine 6-phosphate
-
?
2.7.1.60 ATP + N-acetyl-D-mannosamine Rattus norvegicus Wistar
-
ADP + N-acetyl-D-mannosamine 6-phosphate
-
?

Organism

EC Number Organism UniProt Comment Textmining
2.7.1.60 Rattus norvegicus
-
Wistar
-
2.7.1.60 Rattus norvegicus Wistar
-
Wistar
-
3.2.1.183 Rattus norvegicus
-
-
-
5.1.3.14 Rattus norvegicus
-
bifunctional enzyme EC 5.1.3.14/EC 2.7.1.60
-

Purification (Commentary)

EC Number Purification (Comment) Organism
2.7.1.60
-
Rattus norvegicus
3.2.1.183 using salmine sulfate precipitation and chromatography on phenyl-Sepharose, ATP-agarose, and Mono Q Rattus norvegicus
5.1.3.14 bifunctional enzyme EC 5.1.3.14/EC 2.7.1.60 Rattus norvegicus

Source Tissue

EC Number Source Tissue Comment Organism Textmining
2.7.1.60 intestinal mucosa
-
Rattus norvegicus
-
2.7.1.60 liver
-
Rattus norvegicus
-
2.7.1.60 salivary gland
-
Rattus norvegicus
-
3.2.1.183 liver
-
Rattus norvegicus
-
5.1.3.14 liver
-
Rattus norvegicus
-

Specific Activity [micromol/min/mg]

EC Number Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
2.7.1.60 4.114
-
-
Rattus norvegicus
5.1.3.14 0.878
-
-
Rattus norvegicus

Storage Stability

EC Number Storage Stability Organism
2.7.1.60 -70°C, no loss of activity observed for several months Rattus norvegicus
5.1.3.14 -70°C, stable for several months Rattus norvegicus

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
2.7.1.60 ATP + N-acetyl-D-mannosamine
-
Rattus norvegicus ADP + N-acetyl-D-mannosamine 6-phosphate
-
?
2.7.1.60 ATP + N-acetyl-D-mannosamine
-
Rattus norvegicus Wistar ADP + N-acetyl-D-mannosamine 6-phosphate
-
?
2.7.1.60 ATP + N-acetyl-D-mannosamine
-
Rattus norvegicus ADP + N-acetyl-D-mannosamine-6-phosphate
-
?
2.7.1.60 ATP + N-acetyl-D-mannosamine
-
Rattus norvegicus Wistar ADP + N-acetyl-D-mannosamine-6-phosphate
-
?
2.7.1.60 additional information UDP-GlcNAc 2-epimerase and ManNAc kinase are parts of one bifunctional enzyme, catalyzes the first 2 steps in N-acetylneuraminic acid biosynthesis Rattus norvegicus ?
-
?
2.7.1.60 additional information UDP-GlcNAc 2-epimerase and ManNAc kinase are parts of one bifunctional enzyme, catalyzes the first 2 steps in N-acetylneuraminic acid biosynthesis Rattus norvegicus Wistar ?
-
?
3.2.1.183 UDP-N-acetyl-alpha-D-glucosamine + H2O
-
Rattus norvegicus N-acetyl-alpha-D-mannosamine + UDP
-
?
3.2.1.183 UDP-N-acetyl-alpha-D-glucosamine + H2O
-
Rattus norvegicus Wistar N-acetyl-alpha-D-mannosamine + UDP
-
?

Subunits

EC Number Subunits Comment Organism
2.7.1.60 dimer 2 * 75000, self associates as a dimer, gel filtration, SDS-PAGE, catalyzes only phosphorylation of N-acetylmannosamine, incubation with UDP-N-acetylglucosamine leads to reassembly of the fully active hexamer Rattus norvegicus
2.7.1.60 hexamer 6 * 75000, self associates as a hexamer, gel filtration, SDS-PAGE, catalyzes both enzyme activities, UDP-GlcNAc-epimerase/ManNAc kinase Rattus norvegicus
3.2.1.183 dimer 2 * 75000, gel filtration Rattus norvegicus
3.2.1.183 hexamer 6 * 75000, gel filtration Rattus norvegicus
5.1.3.14 hexamer bifunctional enzyme EC 5.1.3.14/EC 2.7.1.60, SDS-PAGE Rattus norvegicus

Synonyms

EC Number Synonyms Comment Organism
2.7.1.60 ManAc kinase
-
Rattus norvegicus
2.7.1.60 UDP-GlcNAc-epimerase/ManNAc kinase
-
Rattus norvegicus
3.2.1.183 UDP-GlcNAc 2-epimerase bifunctional enzyme complex consisting of UDP-N-acetylglucosamine 2-epimerase and and N-acetylmannosamine kinase Rattus norvegicus
3.2.1.183 UDP-N-acetylglucosamine 2-epimerase
-
Rattus norvegicus

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
3.2.1.183 37
-
assay at Rattus norvegicus

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
3.2.1.183 7.5
-
assay at Rattus norvegicus