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Literature summary for 2.7.7.64 extracted from

  • Yang, T.; Bar-Peled, M.
    Identification of a novel UDP-sugar pyrophosphorylase with a broad substrate specificity in Trypanosoma cruzi (2010), Biochem. J., 429, 533-543.
    View publication on PubMed

Cloned(Commentary)

Cloned (Comment) Organism
expressed in Escherichia coli as a His-tagged fusion protein Trypanosoma cruzi
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 Trypanosoma cruzi

Inhibitors

Inhibitors Comment Organism Structure
EDTA
-
Trypanosoma cruzi

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
additional information
-
additional information kinetics, overview Trypanosoma cruzi
0.026
-
UDP-D-glucose pH 7.6, 37°C, recombinant enzyme Trypanosoma cruzi
0.026
-
UDP-glucose reverse reaction, pH 7.6, 37°C Trypanosoma cruzi
0.0284
-
UTP pH 7.6, 37°C, recombinant enzyme Trypanosoma cruzi
0.0284
-
UTP forward reaction, pH 7.6, 37°C Trypanosoma cruzi
0.134
-
diphosphate reverse reaction, pH 7.6, 37°C Trypanosoma cruzi
0.136
-
diphosphate pH 7.6, 37°C, recombinant enzyme Trypanosoma cruzi
0.177
-
alpha-D-glucose 1-phosphate pH 7.6, 37°C, recombinant enzyme Trypanosoma cruzi
0.177
-
D-glucose 1-phosphate co-substrate: UTP, pH 7.6, 37°C Trypanosoma cruzi
0.177
-
D-glucose 1-phosphate forward reaction, pH 7.6, 37°C Trypanosoma cruzi
0.818
-
D-galactose 1-phosphate co-substrate: UTP, pH 7.6, 37°C Trypanosoma cruzi
1.76
-
D-glucose 1-phosphate co-substrate: TTP, pH 7.6, 37°C Trypanosoma cruzi
2.538
-
D-xylose 1-phosphate co-substrate: UTP, pH 7.6, 37°C Trypanosoma cruzi
2.54
-
TTP co-substrate: glucose 1-phosphate, pH 7.6, 37°C Trypanosoma cruzi

Metals/Ions

Metals/Ions Comment Organism Structure
Mg2+ Mg2+ or Mn2+ required Trypanosoma cruzi
Mg2+ the enzyme requires bivalent ions, Mg2+ or Mn2+, for its activity Trypanosoma cruzi
Mn2+ Mg2+ or Mn2+ required Trypanosoma cruzi
Mn2+ the enzyme requires bivalent ions, Mg2+ or Mn2+, for its activity Trypanosoma cruzi

Molecular Weight [Da]

Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
67000
-
SDS-PAGE Trypanosoma cruzi
67000
-
x * 67000, recombinant His-tagged enzyme, SDS-PAGE Trypanosoma cruzi

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
additional information Trypanosoma cruzi the UDP-sugar pyrophosphorylase of Trypanosoma cruzi shows a broader substrate specificity, the nucleotidyltransferase is capable of transforming different types of sugar 1-phosphates and NTP into NDP-sugars ?
-
?
TTP + alpha-D-glucose 1-phosphate Trypanosoma cruzi
-
diphosphate + TDP-D-glucose
-
?
UTP + alpha-D-galactose 1-phosphate Trypanosoma cruzi
-
diphosphate + UDP-D-galactose
-
?
UTP + alpha-D-glucose 1-phosphate Trypanosoma cruzi
-
diphosphate + UDP-D-glucose
-
r
UTP + alpha-D-glucuronate 1-phosphate Trypanosoma cruzi
-
diphosphate + UDP-D-glucuronate
-
?
UTP + alpha-D-xylose 1-phosphate Trypanosoma cruzi
-
diphosphate + UDP-D-xylose
-
?

Organism

Organism UniProt Comment Textmining
Trypanosoma cruzi
-
-
-
Trypanosoma cruzi D3YBH2 gene Sloppy
-

Purification (Commentary)

Purification (Comment) Organism
recombinant His-tagged enzyme from Escherichia coli strain BL21(DE3)pLysS by nickel affinity chromatography and gel filtration Trypanosoma cruzi
using Ni-NTA chromatography Trypanosoma cruzi

Storage Stability

Storage Stability Organism
-80°C, flash-frozen in liquid nitrogen, several months Trypanosoma cruzi

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
D-galactose 1-phosphate + UTP
-
Trypanosoma cruzi UDP-galactose + diphosphate
-
r
D-glucose 1-phosphate + TTP
-
Trypanosoma cruzi TTP-glucose + diphosphate
-
r
D-glucose 1-phosphate + UTP
-
Trypanosoma cruzi UDP-glucose + diphosphate
-
r
D-xylose 1-phosphate + UTP
-
Trypanosoma cruzi UDP-xylose + diphosphate
-
r
diphosphate + UDP-D-glucose
-
Trypanosoma cruzi UTP + alpha-D-glucose 1-phosphate
-
r
additional information the UDP-sugar pyrophosphorylase of Trypanosoma cruzi shows a broader substrate specificity, the nucleotidyltransferase is capable of transforming different types of sugar 1-phosphates and NTP into NDP-sugars Trypanosoma cruzi ?
-
?
TTP + alpha-D-glucose 1-phosphate
-
Trypanosoma cruzi diphosphate + TDP-D-glucose
-
?
UTP + alpha-D-galactose 1-phosphate
-
Trypanosoma cruzi diphosphate + UDP-D-galactose
-
?
UTP + alpha-D-glucose 1-phosphate
-
Trypanosoma cruzi diphosphate + UDP-D-glucose
-
r
UTP + alpha-D-glucuronate 1-phosphate
-
Trypanosoma cruzi diphosphate + UDP-D-glucuronate
-
?
UTP + alpha-D-xylose 1-phosphate
-
Trypanosoma cruzi diphosphate + UDP-D-xylose
-
?

Subunits

Subunits Comment Organism
? x * 67000, recombinant His-tagged enzyme, SDS-PAGE Trypanosoma cruzi
More real-time NMR analysis of TcSloppy, overview Trypanosoma cruzi

Synonyms

Synonyms Comment Organism
TcSloppy
-
Trypanosoma cruzi
UDP-sugar pyrophosphorylase
-
Trypanosoma cruzi

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
30 42
-
Trypanosoma cruzi
37
-
assay at Trypanosoma cruzi

Temperature Range [°C]

Temperature Minimum [°C] Temperature Maximum [°C] Comment Organism
30 42 highly active between Trypanosoma cruzi

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
0.085
-
TTP co-substrate: glucose 1-phosphate, pH 7.6, 37°C Trypanosoma cruzi
0.12
-
D-glucose 1-phosphate co-substrate: TTP, pH 7.6, 37°C Trypanosoma cruzi
8.6
-
D-xylose 1-phosphate co-substrate: UTP, pH 7.6, 37°C Trypanosoma cruzi
26
-
D-glucose 1-phosphate co-substrate: UTP, pH 7.6, 37°C Trypanosoma cruzi
35.2
-
D-galactose 1-phosphate co-substrate: UTP, pH 7.6, 37°C Trypanosoma cruzi

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
6
-
maximum activity in phosphate buffer Trypanosoma cruzi
6.5 8
-
Trypanosoma cruzi
7.6
-
assay at Trypanosoma cruzi

pH Range

pH Minimum pH Maximum Comment Organism
6.5 8 highly active between Trypanosoma cruzi
7.5 7.8 maximum activity in Tris buffer Trypanosoma cruzi

General Information

General Information Comment Organism
evolution phylogenetic analysis Trypanosoma cruzi
physiological function the parasite enzyme with such broad substrate specificities suggests an alternative pathway that might play an essential role for nucleotide-sugar biosynthesis and for the regulation of the NDP-sugar pool in the parasite Trypanosoma cruzi

kcat/KM [mM/s]

kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
0.0334
-
TTP co-substrate: glucose 1-phosphate, pH 7.6, 37°C Trypanosoma cruzi
0.068
-
D-glucose 1-phosphate co-substrate: TTP, pH 7.6, 37°C Trypanosoma cruzi
34
-
D-xylose 1-phosphate co-substrate: UTP, pH 7.6, 37°C Trypanosoma cruzi
40
-
D-galactose 1-phosphate co-substrate: UTP, pH 7.6, 37°C Trypanosoma cruzi
150
-
D-glucose 1-phosphate co-substrate: UTP, pH 7.6, 37°C Trypanosoma cruzi
150
-
D-glucose 1-phosphate forward reaction, pH 7.6, 37°C Trypanosoma cruzi
220
-
diphosphate reverse reaction, pH 7.6, 37°C Trypanosoma cruzi
920
-
UTP forward reaction, pH 7.6, 37°C Trypanosoma cruzi
1150
-
UDP-glucose reverse reaction, pH 7.6, 37°C Trypanosoma cruzi