Information on EC 2.7.7.64 - UTP-monosaccharide-1-phosphate uridylyltransferase

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

EC NUMBER
COMMENTARY hide
2.7.7.64
-
RECOMMENDED NAME
GeneOntology No.
UTP-monosaccharide-1-phosphate uridylyltransferase
REACTION
REACTION DIAGRAM
COMMENTARY hide
ORGANISM
UNIPROT
LITERATURE
UTP + a monosaccharide 1-phosphate = diphosphate + UDP-monosaccharide
show the reaction diagram
-
-
-
-
PATHWAY
BRENDA Link
KEGG Link
MetaCyc Link
D-galactose detoxification
-
-
stachyose degradation
-
-
sucrose biosynthesis II
-
-
sucrose degradation II (sucrose synthase)
-
-
type I lipoteichoic acid biosynthesis (S. aureus)
-
-
UDP-alpha-D-glucose biosynthesis I
-
-
Pentose and glucuronate interconversions
-
-
Galactose metabolism
-
-
Ascorbate and aldarate metabolism
-
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Amino sugar and nucleotide sugar metabolism
-
-
Metabolic pathways
-
-
Biosynthesis of antibiotics
-
-
CAS REGISTRY NUMBER
COMMENTARY hide
223918-15-8
-
ORGANISM
COMMENTARY hide
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
group Reticulatus, cultivar Noy Yizre’el
SwissProt
Manually annotated by BRENDA team
L. Merr. cv. Lambert
SwissProt
Manually annotated by BRENDA team
no activity in Homo sapiens
-
-
-
Manually annotated by BRENDA team
-
-
-
Manually annotated by BRENDA team
GK24
-
-
Manually annotated by BRENDA team
GENERAL INFORMATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
evolution
phylogenetic analysis
malfunction
-
A knock-out of the USP gene results in non-fertile pollen. Mutant plants show an arabinose reduction in the cell wall, and accumulate mainly two sugars, arabinose and xylose, in the cytoplasm
metabolism
-
the enzyme, likely involved in monosaccharide salvage, preferentially generates UDP-glucose and UDP-galactose, but it may also activate other hexose- or pentose-1-phosphates such as galacturonic acid-1-phosphate or arabinose-1-phosphate
physiological function
additional information
-
active site conformations of apoenzyme and ligand-bound enzyme, substrate binding structures, structure-function analysis, modeling, overview
SUBSTRATE
PRODUCT                       
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
4-azido-D-galactose 1-phosphate + UTP
UDP-4-azido-D-galactose + diphosphate
show the reaction diagram
-
-
43% conversion rate
-
?
6-deoxy-D-galactose 1-phosphate + UTP
UDP-6-deoxy-D-galactose + diphosphate
show the reaction diagram
-
-
29% conversion rate
-
?
D-galactose 1-phosphate + dTTP
dTDP-D-galactose + diphosphate
show the reaction diagram
-
-
-
-
?
D-galactose 1-phosphate + dUTP
dUDP-D-galactose + diphosphate
show the reaction diagram
-
-
-
-
?
D-galactose 1-phosphate + TTP
TDP-D-galactose + diphosphate
show the reaction diagram
-
-
-
?
D-galactose 1-phosphate + UTP
UDP-D-galactose + diphosphate
show the reaction diagram
D-galactose 1-phosphate + UTP
UDP-galactose + diphosphate
show the reaction diagram
D-galacturonate 1-phosphate + UTP
UDP-D-galacturonate + diphosphate
show the reaction diagram
-
-
-
?
D-glucose 1-phosphate + TTP
TTP-glucose + diphosphate
show the reaction diagram
-
-
-
r
D-glucose 1-phosphate + UTP
UDP-D-glucose + diphosphate
show the reaction diagram
D-glucose 1-phosphate + UTP
UDP-glucose + diphosphate
show the reaction diagram
D-glucuronic acid 1-phosphate + UTP
UDP-glucuronic acid + diphosphate
show the reaction diagram
-
-
-
?
D-xylose 1-phosphate + UTP
UDP-xylose + diphosphate
show the reaction diagram
diphosphate + UDP-D-glucose
UTP + alpha-D-glucose 1-phosphate
show the reaction diagram
-
-
-
r
diphosphate + UDP-D-glucose
UTP + D-glucose 1-phosphate
show the reaction diagram
-
-
-
r
L-arabinopyranose 1-phosphate + UTP
UDP-L-arabinopyranose + diphosphate
show the reaction diagram
-
-
-
?
L-arabinose 1-phosphate + UTP
UDP-L-arabinose + diphosphate
show the reaction diagram
-
-
-
?
N-acetylglucosamine 1-phosphate + UTP
UDP-N-acetylglucosamine + diphosphate
show the reaction diagram
poorly catalysed reaction
-
-
?
TTP + alpha-D-glucose 1-phosphate
diphosphate + TDP-D-glucose
show the reaction diagram
-
-
-
?
UDP + alpha-D-glucose 1-phosphate
UDP-alpha-D-glucose + diphosphate
show the reaction diagram
UDP-galacturonic acid + diphosphate
galacturonic acid 1-phosphate + UTP
show the reaction diagram
-
reduced activity in floret extracts depleted for AtUSP by immunprecipitation: only endogenous UDP-galacturonic acid-utilizing pyrophosphorylase activity and final step in myo-inositol oxidation pathway
-
-
?
UDP-monosaccharide + diphosphate
monosaccharide 1-phosphate + UTP
show the reaction diagram
salvage pathway
-
-
r
UTP + a monosaccharide 1-phosphate
diphosphate + UDP-monosaccharide
show the reaction diagram
the enzyme catalyzes the formation of various UDP-sugars at the end of salvage pathways
-
-
?
UTP + alpha-D-galactose 1-phosphate
diphosphate + UDP-D-galactose
show the reaction diagram
UTP + alpha-D-galactose 1-phosphate
diphosphate + UDP-galactose
show the reaction diagram
-
-
-
r
UTP + alpha-D-galacturonate
diphosphate + UDP-alpha-D-galacturonate
show the reaction diagram
-
-
-
r
UTP + alpha-D-galacturonate 1-phosphate
diphosphate + UDP-D-galacturonate
show the reaction diagram
-
-
-
-
?
UTP + alpha-D-glucose 1-phosphate
diphosphate + UDP-D-glucose
show the reaction diagram
UTP + alpha-D-glucose 1-phosphate
diphosphate + UDP-glucose
show the reaction diagram
-
-
-
r
UTP + alpha-D-glucuronate 1-phosphate
diphosphate + UDP-D-glucuronate
show the reaction diagram
-
-
-
?
UTP + alpha-D-xylose 1-phosphate
diphosphate + UDP-D-xylose
show the reaction diagram
-
-
-
?
UTP + alpha-L-arabinose 1-phosphate
diphosphate + UDP-L-arabinose
show the reaction diagram
-
-
-
-
?
UTP + arabinose 1-phosphate
diphosphate + UDP-arabinose
show the reaction diagram
70.5% of enzyme activity with glucose 1-phosphate
-
-
r
UTP + beta-L-arabinopyranose
diphosphate + UDP-beta-L-arabinopyranose
show the reaction diagram
-
-
-
r
UTP + D-galactose 1-phosphate
diphosphate + UDP-galactose
show the reaction diagram
UTP + D-galacturonic acid 1-phosphate
diphosphate + UDP-galacturonic acid
show the reaction diagram
-
equilibrium constant is 0.24
-
-
r
UTP + D-glucose 1-phosphate
diphosphate + UDP-glucose
show the reaction diagram
UTP + D-glucuronic acid 1-phosphate
diphosphate + UDP-D-glucuronic acid
show the reaction diagram
71.3% of enzyme activity with glucose 1-phosphate
-
-
r
UTP + D-glucuronic acid 1-phosphate
diphosphate + UDP-glucuronic acid
show the reaction diagram
UTP + D-mannose 1-phosphate
diphosphate + UDP-mannose
show the reaction diagram
UTP + D-xylopyranose 1-phosphate
diphosphate + UDP-D-xylopyranose
show the reaction diagram
-
-
-
r
UTP + D-xylose 1-phosphate
diphosphate + UDP-alpha-D-xylopyranose
show the reaction diagram
UTP + D-xylose 1-phosphate
diphosphate + UDP-xylose
show the reaction diagram
35.9% of enzyme activity with glucose 1-phosphate
-
-
r
UTP + L-arabinose 1-phosphate
diphosphate + UDP-beta-L-arabinopyranose
show the reaction diagram
-
-
-
?
UTP + monosaccharide 1-phosphate
diphosphate + UDP-monosaccharide
show the reaction diagram
-
the enzyme has a critical role in pollen development. The products of the AtUSP reaction can act as precursors for the synthesis of glycolipids, glycoproteins, and cell wall components including pectin and hemicellulose
-
-
?
UTP + N-acetyl-D-glucosamine 1-phosphate
diphosphate + UDP-N-acetylglucosamine
show the reaction diagram
4.3% of enzyme activity with glucose 1-phosphate
-
-
r
UTP + N-acetylglucosamine 1-phosphate
diphosphate + UDP-N-acetylglucosamine
show the reaction diagram
additional information
?
-
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
TTP + alpha-D-glucose 1-phosphate
diphosphate + TDP-D-glucose
show the reaction diagram
D3YBH2
-
-
-
?
UDP-galacturonic acid + diphosphate
galacturonic acid 1-phosphate + UTP
show the reaction diagram
-
reduced activity in floret extracts depleted for AtUSP by immunprecipitation: only endogenous UDP-galacturonic acid-utilizing pyrophosphorylase activity and final step in myo-inositol oxidation pathway
-
-
?
UDP-monosaccharide + diphosphate
monosaccharide 1-phosphate + UTP
show the reaction diagram
Q9C5I1
salvage pathway
-
-
r
UTP + a monosaccharide 1-phosphate
diphosphate + UDP-monosaccharide
show the reaction diagram
Q5W915
the enzyme catalyzes the formation of various UDP-sugars at the end of salvage pathways
-
-
?
UTP + alpha-D-galactose 1-phosphate
diphosphate + UDP-D-galactose
show the reaction diagram
D3YBH2
-
-
-
?
UTP + alpha-D-glucose 1-phosphate
diphosphate + UDP-D-glucose
show the reaction diagram
D3YBH2
-
-
-
r
UTP + alpha-D-glucuronate 1-phosphate
diphosphate + UDP-D-glucuronate
show the reaction diagram
D3YBH2
-
-
-
?
UTP + alpha-D-xylose 1-phosphate
diphosphate + UDP-D-xylose
show the reaction diagram
D3YBH2
-
-
-
?
UTP + D-galactose 1-phosphate
diphosphate + UDP-galactose
show the reaction diagram
D3G6S4, Q4QE75
-
-
-
r
UTP + D-glucose 1-phosphate
diphosphate + UDP-glucose
show the reaction diagram
D3G6S4, Q4QE75
-
-
-
r
UTP + monosaccharide 1-phosphate
diphosphate + UDP-monosaccharide
show the reaction diagram
-
the enzyme has a critical role in pollen development. The products of the AtUSP reaction can act as precursors for the synthesis of glycolipids, glycoproteins, and cell wall components including pectin and hemicellulose
-
-
?
additional information
?
-
METALS and IONS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
Co2+
-
divalent metal required. Mn2+ (3 mM) is 63% as effective as Mg2+ (3 mM)
INHIBITORS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
hydrogen peroxide
inactivation, reactivation by reduction using cysteine or thioredoxin
nitric oxide
inactivation, reactivation by reduction using cysteine or thioredoxin
ACTIVATING COMPOUND
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
fructose 1-phosphate
-
2 mM, 78% activation
galactose 6-phosphate
-
2 mM, 70% activation
KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.43
alpha-D-galactose 1-phosphate
-
0.177 - 0.27
alpha-D-glucose 1-phosphate
0.96
arabinose 1-phosphate
pH 7.0, 35°C
0.09 - 0.86
D-galactose 1-phosphate
1.15
D-galacturonic acid 1-phosphate
-
pH 7.0, 35°C
0.13 - 1.76
D-glucose 1-phosphate
0.094 - 0.48
D-glucuronic acid 1-phosphate
1 - 2.538
D-xylose 1-phosphate
0.134 - 1.018
diphosphate
8.14
dTTP
-
pH not specified in the publication, temperature not specified in the publication
3.74
dUTP
-
pH not specified in the publication, temperature not specified in the publication
0.96 - 1.5
L-arabinose 1-phosphate
0.6
N-acetylglucosamine 1-phosphate
-
pH 8.0, 73°C
0.9 - 2.54
TTP
0.79
UDP-alpha-D-galacturonate
reverse reaction, pH 7.8, 25°C
0.373
UDP-beta-L-arabinopyranose
reverse reaction, pH 7.8, 25°C
0.148
UDP-D-galactose
reverse reaction, pH 7.8, 25°C
0.79
UDP-D-galacturonate
reverse reaction, pH 7.8, 25°C
0.026 - 0.34
UDP-D-glucose
0.148 - 0.44
UDP-galactose
1.26
UDP-galacturonic acid
-
pH 7.0, 35°C
0.026 - 0.34
UDP-glucose
0.373
UDP-L-arabinopyranose
reverse reaction, pH 7.8, 25°C
0.0284 - 2.88
UTP
additional information
additional information
-
TURNOVER NUMBER [1/s]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
35.2 - 219
D-galactose 1-phosphate
0.008 - 191
D-glucose 1-phosphate
67
D-glucuronic acid 1-phosphate
-
pH 7, 35°C
8.6 - 51
D-xylose 1-phosphate
15 - 185
diphosphate
27
dTTP
-
pH not specified in the publication, temperature not specified in the publication
219
dUTP
-
pH not specified in the publication, temperature not specified in the publication
138
L-arabinose 1-phosphate
-
pH 7, 35°C
0.2
N-acetylglucosamine 1-phosphate
-
pH 8.0, 73°C
0.085
TTP
co-substrate: glucose 1-phosphate, pH 7.6, 37°C
13
UDP-alpha-D-galacturonate
reverse reaction, pH 7.8, 25°C
72
UDP-beta-L-arabinopyranose
reverse reaction, pH 7.8, 25°C
154
UDP-D-galactose
reverse reaction, pH 7.8, 25°C
13
UDP-D-galacturonate
reverse reaction, pH 7.8, 25°C
180
UDP-D-glucose
reverse reaction, pH 7.8, 25°C
154
UDP-galactose
reverse reaction, pH 7.8, 25°C
133 - 180
UDP-glucose
72
UDP-L-arabinopyranose
reverse reaction, pH 7.8, 25°C
0.003 - 348
UTP
additional information
D-galactose 1-phosphate
kcat/KM VALUE [1/mMs-1]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
40 - 259
D-galactose 1-phosphate
0.0054 - 150
D-glucose 1-phosphate
34
D-xylose 1-phosphate
co-substrate: UTP, pH 7.6, 37°C
20.8 - 580
diphosphate
3.3
dTTP
-
pH not specified in the publication, temperature not specified in the publication
59
dUTP
-
pH not specified in the publication, temperature not specified in the publication
0.0334
TTP
co-substrate: glucose 1-phosphate, pH 7.6, 37°C
1070
UDP-D-galactose
reverse reaction, pH 7.8, 25°C
16.7
UDP-D-galacturonate
reverse reaction, pH 7.8, 25°C
1060
UDP-D-glucose
reverse reaction, pH 7.8, 25°C
1150
UDP-glucose
reverse reaction, pH 7.6, 37°C
198
UDP-L-arabinopyranose
reverse reaction, pH 7.8, 25°C
0.0015 - 2090
UTP
SPECIFIC ACTIVITY [µmol/min/mg]
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
1.2
-
+/-0.08 micromol/min/mg, N-acetylglucosamine 1-phosphate
22.8
-
+/-1.1 micromol/min/mg, D-xylose 1-phosphate
30
UDP-L-arabinose pyrophosphorylase, pH 7.0, 35°C
44.5
-
+/-0.9 micromol/min/mg, L-arabinose 1-phosphate
51.4
-
+/-3.6 micromol/min/mg, D-glucuronic acid 1-phosphate
63.9
; UDP-D-glucose pyrophosphorylase, pH 7.0, 35°C
84.3
-
+/-2.4 micromol/min/mg, D-glucose 1-phosphate
86.9
-
+/-2.4 micromol/min/mg, D-galactose 1-phosphate
95
recombinant UDP-glucose pyrophosphorylase, pH 7.0, 35°C
223
with UDP-glucose as a substrate
233
with alpha-D-galactose 1-phosphate as a substrate
410
with alpha-D-glucose 1-phosphate as a substrate
678
with UDP-galactose as a substrate
additional information
pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
6.9
-
in presence of Mg2+
7.8
activity assay; assay at
pH RANGE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
5 - 9
-
pH 5.0: about 75% of maximal activity, pH 9.0: about 50% of maximal activity
5.5 - 8
-
pH 5.5: about 90% of maximal activity, pH 8.0: about 75% of maximal activity
6 - 8
-
60% of maximum UDP-glucose-synthesising activity
6.5 - 8
highly active between
7.5 - 7.8
maximum activity in Tris buffer
TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
25
activity assay; assay at
TEMPERATURE RANGE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
30 - 42
highly active between
65 - 95
-
65°C: about 60% of maximal activity, 95°C: 90% of maximal activity
pI VALUE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
5.82
calculated
6.1
-
theoretical
SOURCE TISSUE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
SOURCE
-
enzyme assay on homogenate of transgenic plants
Manually annotated by BRENDA team
-
AtUSP mRNA-specific qRT-PCR
Manually annotated by BRENDA team
-
AtUSP mRNA-specific qRT-PCR, strong expression in vascular tissue (promoter: beta-glucuronidase assay)
Manually annotated by BRENDA team
-
relatively high levels in most cell types (AtUSP mRNA-specific qRT-PCR)
Manually annotated by BRENDA team
-
young leaves (AtUSP mRNA-specific qRT-PCR), strong expression in vascular tissue (promoter: beta-glucuronidase assay)
Manually annotated by BRENDA team
-
33-days-old
Manually annotated by BRENDA team
-
AtUSP mRNA-specific qRT-PCR
Manually annotated by BRENDA team
LOCALIZATION
ORGANISM
UNIPROT
COMMENTARY hide
GeneOntology No.
LITERATURE
SOURCE
-
USP1 and USP2, differential centrifugation and Western blot
Manually annotated by BRENDA team
-
USP1, differential centrifugation and Western blot
-
Manually annotated by BRENDA team
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
41000
-
1 * 41000, SDS-PAGE
45000
-
gel filtration
50000
gel filtration
66040
x * 66040, calculated from sequence
67850
-
deduced from amino acid sequence
68000
SDS-PAGE
69000
predicted from cDNA
69700
USP-His6, apparent molecular mass determind by gel filtration and standard kurve
70400
theoretical
SUBUNITS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
monomer
additional information
Crystallization/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
crystal structures of a USP in the APO-, UTP-, and UDP-sugar-bound forms is presented. The overall structure of the enzyme exhibits a significant structural homology to other nucleotidyldiphosphate-glucose pyrophosphorylases. The broad substrate specificity is strongly correlated with the flexibility of the SB-loop and, in contrast to UDP-glucose pyrophosphorylase of Leishmania major, the spacious sugar binding cavity as well as the multiple conformations observed for the residues involved in sugar moiety binding
-
pH STABILITY
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
6.5 - 8
80% loss of activity below pH 5.0
669277
TEMPERATURE STABILITY
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
55
-
loss of UDP-glucose-synthesising activity
95
-
half-life: 30 min
additional information
GENERAL STABILITY
ORGANISM
UNIPROT
LITERATURE
USP2 is not product of proteolytic cleavage of USP1
-
STORAGE STABILITY
ORGANISM
UNIPROT
LITERATURE
-80°C, flash-frozen in liquid nitrogen, several months
Purification/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
ammonium sulfate fractionation, ion exchange chromatography (DEAE), hydrophobic interaction chromatography, gel filtration and ion exchange chromatography (DEAE), recombinant protein: immobilized metal ion affinity chromatography, splitting off the fused thioredoxin and His-tag, ion exchange chromatography (DEAE); recombinant
chelating chromatography (elution: 250 mM imidazole) followed by cleavage of N-terminal thioredoxin- and His-tag with thrombin and DEAE Sepharose FF chromatography
-
MonoQ HR 5/5 column chromatography
on a 1-ml HisTrap HP Ni2+-chelating, a HiPrep 26/10 desalting, and a 1-ml Q-Sepharose FF anion exchange column; using Ni-NTA chromatography, ion-exchange and gel filtration
recombinant His-tagged enzyme from Escherichia coli strain BL21(DE3)pLysS by nickel affinity chromatography and gel filtration; using Ni-NTA chromatography
using Ni-NTA chromatography
-
Cloned/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
; His-tagged thioredoxin fusion protein expressed in Escherichia coli BL21gold strain
expressed as a His-tagged fusion protein
-
expressed in Escherichia coli
expressed in Escherichia coli as a His-tagged fusion protein; gene TcSloppy, DNA and amino acid sequence determination and analysis, phylogenetic analysis and sequence comparisons, expression of His-tagged enzyme in Escherichia coli strain BL21(DE3)pLysS
expressed in Escherichia coli as a His-tagged fusion protein; into the vector pET22b for expression in Escherichia coli BL21DE3 cells
expressed in Escherichia coli BL21(DE3) cells
-
expression in Agrobacterium tumefaciens
expression in Escherichia coli
from cDNA generated from total RNA from 2-week-old seedlings in pGEM T-Easy for amplification and in pET32a for inducible expression with N-terminal thioredoxin- and His-tag in Escherichia coli BL21(DE3) and in pBI121 for generation of transgenic plants via Agrobacterium tumefaciens
-
APPLICATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
molecular biology
-
polyclonal rabbit anti-AtUSP antibody recognizes USP1 and USP2
synthesis