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Literature summary for 2.7.7.64 extracted from

  • Kotake, T.; Hojo, S.; Yamaguchi, D.; Aohara, T.; Konishi, T.; Tsumuraya, Y.
    Properties and physiological functions of UDP-sugar pyrophosphorylase in Arabidopsis (2007), Biosci. Biotechnol. Biochem., 71, 761-771.
    View publication on PubMed
Search for more literature for 2.7.7.64

Cloned(Commentary)

Cloned (Comment) Organism
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 Arabidopsis thaliana

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.077
-
 UTP pH 7, 35°C Arabidopsis thaliana
0.094
-
 D-glucuronic acid 1-phosphate pH 7, 35°C Arabidopsis thaliana
0.16
-
 diphosphate pH 7, 35°C Arabidopsis thaliana
0.23
-
 D-glucose 1-phosphate pH 7, 35°C Arabidopsis thaliana
0.27
-
 D-galactose 1-phosphate pH 7, 35°C Arabidopsis thaliana
0.28
-
 UDP-glucose pH 7, 35°C Arabidopsis thaliana
1
-
 D-xylose 1-phosphate pH 7, 35°C Arabidopsis thaliana
1.5
-
 L-arabinose 1-phosphate pH 7, 35°C Arabidopsis thaliana

Metals/Ions

Metals/Ions Comment Organism Structure
Mg2+ no activity in absence of divalent cations Arabidopsis thaliana
Mn2+ no activity in absence of divalent cations Arabidopsis thaliana

Molecular Weight [Da]

Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
67850
-
 deduced from amino acid sequence Arabidopsis thaliana
70000
-
 purified recombinant enzyme, SDS-PAGE Arabidopsis thaliana

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
UDP-monosaccharide + diphosphate Arabidopsis thaliana salvage pathway monosaccharide 1-phosphate + UTP
-
 r

Organism

Organism UniProt Comment Textmining
Arabidopsis thaliana Q9C5I1 ecotype Columbia, Landsberg erecta, Wassilewskija
-

Purification (Commentary)

Purification (Comment) Organism
chelating chromatography (elution: 250 mM imidazole) followed by cleavage of N-terminal thioredoxin- and His-tag with thrombin and DEAE Sepharose FF chromatography Arabidopsis thaliana

Source Tissue

Source Tissue Comment Organism Textmining
aerial part enzyme assay on homogenate of transgenic plants Arabidopsis thaliana
-
cauline leaf AtUSP mRNA-specific qRT-PCR Arabidopsis thaliana
-
cotyledon AtUSP mRNA-specific qRT-PCR, strong expression in vascular tissue (promoter: beta-glucuronidase assay) Arabidopsis thaliana
-
flower relatively high levels in most cell types (AtUSP mRNA-specific qRT-PCR) Arabidopsis thaliana
-
inflorescence stem AtUSP mRNA-specific qRT-PCR Arabidopsis thaliana
-
leaf young leaves (AtUSP mRNA-specific qRT-PCR), strong expression in vascular tissue (promoter: beta-glucuronidase assay) Arabidopsis thaliana
-
pollen AtUSP mRNA-specific qRT-PCR, strong expression in vascular tissue (promoter: beta-glucuronidase assay), essential for pollen development since loss-of-function mutation in enzyme causes male infertility Arabidopsis thaliana
-
root strong expression in growing region (promoter: beta-glucuronidase assay), young roots (AtUSP mRNA-specific qRT-PCR) Arabidopsis thaliana
-
rosette leaf AtUSP mRNA-specific qRT-PCR Arabidopsis thaliana
-
seedling young seedlings, expression in most cell types (AtUSP mRNA-specific qRT-PCR) Arabidopsis thaliana
-

Specific Activity [micromol/min/mg]

Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
additional information
-
 143-255% UDP-L-arabinose synthesising activity in extracts from transgenic plants (aerial parts) compared to wild-type plant Arabidopsis thaliana
additional information
-
 25% of wild-type UDP-L-arabinose synthesising activity in plant extracts (aerial parts) upon repression of AtUSP gene expression Arabidopsis thaliana
additional information
-
 no activity detectable for substrates L-fucose 1-phosphate and mannose 1-phosphate Arabidopsis thaliana
1.2
-
 +/-0.08 micromol/min/mg, N-acetylglucosamine 1-phosphate Arabidopsis thaliana
22.8
-
 +/-1.1 micromol/min/mg, D-xylose 1-phosphate Arabidopsis thaliana
44.5
-
 +/-0.9 micromol/min/mg, L-arabinose 1-phosphate Arabidopsis thaliana
51.4
-
 +/-3.6 micromol/min/mg, D-glucuronic acid 1-phosphate Arabidopsis thaliana
84.3
-
 +/-2.4 micromol/min/mg, D-glucose 1-phosphate Arabidopsis thaliana
86.9
-
 +/-2.4 micromol/min/mg, D-galactose 1-phosphate Arabidopsis thaliana

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
D-galactose 1-phosphate + UTP
-
 Arabidopsis thaliana UDP-galactose + diphosphate
-
 ?
D-glucose 1-phosphate + UTP pH 7, 35°C, 2 mM Mg2+ Arabidopsis thaliana UDP-glucose + diphosphate reaction stop by boiling, analysis of products by HPLC (monitoring at 262 nm) r
D-glucuronic acid 1-phosphate + UTP
-
 Arabidopsis thaliana UDP-glucuronic acid + diphosphate
-
 ?
D-xylose 1-phosphate + UTP
-
 Arabidopsis thaliana UDP-xylose + diphosphate
-
 ?
L-arabinose 1-phosphate + UTP
-
 Arabidopsis thaliana UDP-L-arabinose + diphosphate
-
 ?
additional information no activity detectable for substrates L-fucose 1-phosphate and mannose 1-phosphate Arabidopsis thaliana ?
-
 ?
N-acetylglucosamine 1-phosphate + UTP poorly catalysed reaction Arabidopsis thaliana UDP-N-acetylglucosamine + diphosphate
-
 ?
UDP-monosaccharide + diphosphate salvage pathway Arabidopsis thaliana monosaccharide 1-phosphate + UTP
-
 r

Synonyms

Synonyms Comment Organism
AtUSP
-
 Arabidopsis thaliana
UDP-sugar pyrophosphorylase
-
 Arabidopsis thaliana
USP
-
 Arabidopsis thaliana

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
45
-
 UDP-glucose-synthesising activity Arabidopsis thaliana

Temperature Stability [°C]

Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
55
-
 loss of UDP-glucose-synthesising activity Arabidopsis thaliana

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
additional information
-
 UTP Kcat/KM: 1400000 1/M/sec, pH 7, 35°C Arabidopsis thaliana
additional information
-
 UDP-glucose Kcat/KM: 480000 1/M/sec, pH 7, 35°C Arabidopsis thaliana
additional information
-
 D-galactose 1-phosphate Kcat/KM: 490000 1/M/sec, pH 7, 35°C Arabidopsis thaliana
additional information
-
 D-xylose 1-phosphate Kcat/KM: 51000 1/M/sec, pH 7, 35°C Arabidopsis thaliana
additional information
-
 D-glucose 1-phosphate Kcat/KM: 530000 1/M/sec, pH 7, 35°C Arabidopsis thaliana
additional information
-
 D-glucuronic acid 1-phosphate Kcat/KM: 710000 1/M/sec, pH 7, 35°C Arabidopsis thaliana
additional information
-
 diphosphate Kcat/KM: 860000 1/M/sec, pH 7, 35°C Arabidopsis thaliana
additional information
-
 L-arabinose 1-phosphate Kcat/KM: 92000 1/M/sec, pH 7, 35°C Arabidopsis thaliana
51
-
 D-xylose 1-phosphate pH 7, 35°C Arabidopsis thaliana
67
-
 D-glucuronic acid 1-phosphate pH 7, 35°C Arabidopsis thaliana
108
-
 UTP pH 7, 35°C Arabidopsis thaliana
122
-
 D-glucose 1-phosphate pH 7, 35°C Arabidopsis thaliana
133
-
 UDP-glucose pH 7, 35°C Arabidopsis thaliana
133
-
 D-galactose 1-phosphate pH 7, 35°C Arabidopsis thaliana
138
-
 diphosphate pH 7, 35°C Arabidopsis thaliana
138
-
 L-arabinose 1-phosphate pH 7, 35°C Arabidopsis thaliana

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
7
-
 UDP-glucose-synthesising activity Arabidopsis thaliana

pH Range

pH Minimum pH Maximum Comment Organism
6 8 60% of maximum UDP-glucose-synthesising activity Arabidopsis thaliana

pI Value

Organism Comment pI Value Maximum pI Value
Arabidopsis thaliana theoretical
-
 6.1