Information on EC 2.7.7.83 - UDP-N-acetylgalactosamine diphosphorylase

for references in articles please use BRENDA:EC2.7.7.83
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The expected taxonomic range for this enzyme is: Eukaryota, Archaea

EC NUMBER
COMMENTARY hide
2.7.7.83
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RECOMMENDED NAME
GeneOntology No.
UDP-N-acetylgalactosamine diphosphorylase
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REACTION
REACTION DIAGRAM
COMMENTARY hide
ORGANISM
UNIPROT
LITERATURE
UTP + N-acetyl-alpha-D-galactosamine 1-phosphate = diphosphate + UDP-N-acetyl-alpha-D-galactosamine
show the reaction diagram
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SYSTEMATIC NAME
IUBMB Comments
UTP:N-acetyl-alpha-D-galactosamine-1-phosphate uridylyltransferase
The enzyme from plants and animals also has activity toward N-acetyl-alpha-D-glucosamine 1-phosphate (cf. EC 2.7.7.23, UDP-N-acetylglucosamine diphosphorylase) [1,2].
ORGANISM
COMMENTARY hide
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
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SwissProt
Manually annotated by BRENDA team
Sulfolobus tokodaii DSM 16993 / JCM 10545 / NBRC 100140 / 7
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SwissProt
Manually annotated by BRENDA team
GENERAL INFORMATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
physiological function
additional information
SUBSTRATE
PRODUCT                       
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
diphosphate + UDP-N-acetyl-alpha-D-galactosamine
UTP + N-acetyl-alpha-D-galactosamine 1-phosphate
show the reaction diagram
diphosphate + UDP-N-acetylglucosamine
UTP + N-acetylglucosamine 1-phosphate
show the reaction diagram
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r
UTP + N-acetyl-alpha-D-galactosamine 1-phosphate
diphosphate + UDP-N-acetyl-alpha-D-galactosamine
show the reaction diagram
UTP + N-acetyl-D-galactosamine 1-phosphate
diphosphate + UDP-N-acetyl-D-galactosamine
show the reaction diagram
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r
UTP + N-acetyl-D-glucosamine 1-phosphate
diphosphate + UDP-N-acetyl-D-glucosamine
show the reaction diagram
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-
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r
UTP + N-acetylglucosamine 1-phosphate
diphosphate + UDP-N-acetylglucosamine
show the reaction diagram
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r
additional information
?
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the enzyme has multiple sugar-1-phosphate nucleotidylyltransferase, EC 2.7.7.37, and amino-sugar-1-phosphate acetyltransferase, EC 2.3.1.157, activities, overview. In addition to glucosamine-1-phosphate acetyltransferase activity, it possesses unique galactosamine-1-phosphate acetyltransferase activity. Also, the enzyme possesses GlcNAc-1-phosphate nucleotidylyltransferase, EC 2.7.7.23, and N-acetyl-D-galactosamine-1-phosphate uridyltransferase, EC 2.7.7.83, activities, as well as the expected glucose-1-phosphate thymidylyltransferase, EC 2.7.7.24, activity
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NATURAL SUBSTRATES
NATURAL PRODUCTS
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
REVERSIBILITY
r=reversible
ir=irreversible
?=not specified
UTP + N-acetyl-alpha-D-galactosamine 1-phosphate
diphosphate + UDP-N-acetyl-alpha-D-galactosamine
show the reaction diagram
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?
METALS and IONS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
Mg2+
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required
KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.38 - 1.3
N-acetyl-D-galactosamine 1-phosphate
0.24 - 0.32
N-acetyl-D-glucosamine 1-phosphate
0.0053 - 0.006
N-acetylglucosamine 1-phosphate
0.5 - 0.53
UDP-N-acetyl-alpha-D-galactosamine
0.032
UDP-N-acetylglucosamine
wild type protein, pH 8.0, 30°C; wild type protein, pH 8.0, 30°C
0.049 - 0.053
UTP
kcat/KM VALUE [1/mMs-1]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
11200 - 12200
N-acetylglucosamine 1-phosphate
180 - 200
UDP-N-acetyl-alpha-D-galactosamine
2000 - 2200
UDP-N-acetylglucosamine
1200 - 1400
UTP
SPECIFIC ACTIVITY [µmol/min/mg]
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
0.119
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GST-fusion protein, N-acetyl-D-galactosamine 1-phosphate, GST protein shows 0.02 micromol/min/mg in the assay, pH 7.5, 37°C
0.3
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GST fusion removed, N-acetyl-D-galactosamine 1-phosphate, pH 7.5, 37°C
0.525
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GST-fusion protein, N-acetyl-D-glucosamine 1-phosphate, GST protein shows 0.033 micromol/min/mg in the assay, pH 7.5, 37°C
0.887
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GST-fusion protein, N-acetyl-D-glucosamine 1-phosphate, GST protein shows 0.033 micromol/min/mg in the assay, pH 7.5, 37°C
0.9
R115A mutant protein, pH 8.0, 30°C
1.342
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GST-fusion protein, N-acetyl-D-galactosamine 1-phosphate, GST protein shows 0.02 micromol/min/mg in the assay, pH 7.5, 37°C
1.5
R115A mutant protein, pH 8.0, 30°C
5.873
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GST fusion removed, N-acetyl-D-glucosamine 1-phosphate, pH 7.5, 37°C
5.919
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GST fusion removed, N-acetyl-D-glucosamine 1-phosphate, pH 7.5, 37°C
14.9
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GST fusion removed, N-acety-D-lgalactosamine 1-phosphate, pH 7.5, 37°C
68
wild type protein, pH 8.0, 30°C
69
wild type protein, pH 8.0, 30°C
pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
SOURCE TISSUE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
SOURCE
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
57000
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x * 57000, SDS-PAGE
64000
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x * 64000, SDS-PAGE
74000
2 * 74000, gel filtration; gel filtration
120000
gel filtration
SUBUNITS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
?
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x * 57000, SDS-PAGE; x * 64000, SDS-PAGE
homodimer
2 * 74000, gel filtration
monomer
74000, gel filtration
trimer
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the C-terminal domain of the ST0452 protein, with its LbetaH structure, appears to be essential for the formation of its trimeric form and, in turn, the high stability of the entire ST0452 protein
additional information
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the enzyme contains the the N-terminal nucleotidylyltransferase domain (residues 1–210) and the C-terminal acetyltransferase domain (residues 211–401), respectively. Comparisons of the crystal structures of the ST0452 protein, PDB ID GGO, and Escherichia coli protein EcGlmU2, PDB ID 2OI5, comparison with ST0452 mutant enzymes, overview. Despite the structural similarities between the N- and C-termini of the ST0452 protein and those of Escherichia coli EcGlmU, the thermostabilities of the two proteins differ greatly, as EcGlmU is a mesophilic enzyme. The structures of these proteins do not correlate directly with their thermostability
Crystallization/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
vapor diffusion technique, PEG600, 20°C, pH5.7; vapor diffusion technique, PEG600, 20°C, pH5.7
Purification/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
affinity chromatography (glutathione-Sepharose), GST part removed
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anion exchange, phenyl interaction and gel filtration; anion exchange, phenyl interaction and gel filtration
Cloned/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
expressed in Escherichia coli XL1 Blue; expressed in Escherichia coli XL1 Blue
gene ST0452, recombinant expression of C-terminally His6-tagged wild-type and mutant enzymes in Escherichia coli strain BL21-Codon Plus(DE3)-RIL
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GST fusion protein expressed in Escherichia coli JM109
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ENGINEERING
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
R115A
activity drastically affected; activity drastically affected, no dimer formation
H308A
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site-directed mutagenesis, the mutant shows highly reduced activity compared to the wild-type
K337A
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site-directed mutagenesis
K340A
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site-directed mutagenesis
N331A
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site-directed mutagenesis
Y311A
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site-directed mutagenesis
additional information
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construction of expression vectors encoding a series of ST0452 C-terminal deletion mutants with hexahistidine tags at their C-termini, designated pST0452(DC005)H, pST0452(DC011)H, pST0452(DC021)H, pST0452(DC031)H, pST0452(DC041) H, pST0452(DC051)H, pST0452(DC071)H, pST0452 (DC121)H and pST0452(DC171)H. The deletion mutants retain the same tertiary structures as the wild-type ST0452 protein