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Literature summary for 2.7.7.64 extracted from

  • Damerow, S.; Lamerz, A.C.; Haselhorst, T.; Fuehring, J.; Zarnovican, P.; von Itzstein, M.; Routier, F.H.
    Leishmania UDP-sugar pyrophosphorylase: the missing link in galactose salvage? (2010), J. Biol. Chem., 285, 878-887.
    View publication on PubMedView publication on EuropePMC
Search for more literature for 2.7.7.64

Cloned(Commentary)

Cloned (Comment) Organism
expressed in Escherichia coli as a His-tagged fusion protein Leishmania major
into the vector pET22b for expression in Escherichia coli BL21DE3 cells Leishmania major

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.098
-
 UTP forward reaction, D-galactose 1-phosphate, pH 7.8, 25°C Leishmania major
0.098
-
 UTP forward reaction, co-substrate: D-galactose 1-phosphate, pH 7.8, 25°C Leishmania major
0.116
-
 UTP forward reaction, D-glucose 1-phosphate, pH 7.8, 25°C Leishmania major
0.116
-
 UTP forward reaction, co-substrate: D-glucose 1-phosphate, pH 7.8, 25°C Leishmania major
0.148
-
 UDP-galactose reverse reaction, pH 7.8, 25°C Leishmania major
0.148
-
 UDP-D-galactose reverse reaction, pH 7.8, 25°C Leishmania major
0.174
-
 UDP-glucose reverse reaction, pH 7.8, 25°C Leishmania major
0.174
-
 UDP-D-glucose reverse reaction, pH 7.8, 25°C Leishmania major
0.307
-
 diphosphate reverse reaction, UDP-galactose, pH 7.8, 25°C Leishmania major
0.307
-
 diphosphate reverse reaction, co-substrate: UDP-D-galactose, pH 7.8, 25°C Leishmania major
0.373
-
 UDP-beta-L-arabinopyranose reverse reaction, pH 7.8, 25°C Leishmania major
0.373
-
 UDP-L-arabinopyranose reverse reaction, pH 7.8, 25°C Leishmania major
0.383
-
 diphosphate reverse reaction, UDP-glucose, pH 7.8, 25°C Leishmania major
0.383
-
 diphosphate reverse reaction, co-substrate: UDP-D-glucose, pH 7.8, 25°C Leishmania major
0.728
-
 diphosphate reverse reaction, UDP-alpha-D-galacturonate, pH 7.8, 25°C Leishmania major
0.728
-
 diphosphate reverse reaction, co-substrate: UDP-D-galacturonate, pH 7.8, 25°C Leishmania major
0.79
-
 UDP-D-galacturonate reverse reaction, pH 7.8, 25°C Leishmania major
0.79
-
 UDP-alpha-D-galacturonate reverse reaction, pH 7.8, 25°C Leishmania major
0.86
-
 D-galactose 1-phosphate forward reaction, pH 7.8, 25°C Leishmania major
1.01
-
 diphosphate reverse reaction, co-substrate: UDP-L-arabinopyranose, pH 7.8, 25°C Leishmania major
1.018
-
 diphosphate reverse reaction, UDP-beta-L-arabinopyranose, pH 7.8, 25°C Leishmania major
1.7
-
 D-glucose 1-phosphate forward reaction, pH 7.8, 25°C Leishmania major
1.706
-
 D-glucose 1-phosphate forward reaction, pH 7.8, 25°C Leishmania major

Molecular Weight [Da]

Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
66000
-
 USP-His6, determined by SDS-PAGE and Western blot analysis Leishmania major
69000
-
 predicted from cDNA Leishmania major
69700
-
 USP-His6, apparent molecular mass determind by gel filtration and standard kurve Leishmania major
70400
-
 theoretical Leishmania major

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
UTP + D-galactose 1-phosphate Leishmania major
-
 diphosphate + UDP-galactose
-
 r
UTP + D-glucose 1-phosphate Leishmania major
-
 diphosphate + UDP-glucose
-
 r

Organism

Organism UniProt Comment Textmining
Leishmania major D3G6S4
-
-
Leishmania major Q4QE75
-
-

Purification (Commentary)

Purification (Comment) Organism
on a 1-ml HisTrap HP Ni2+-chelating, a HiPrep 26/10 desalting, and a 1-ml Q-Sepharose FF anion exchange column Leishmania major
using Ni-NTA chromatography, ion-exchange and gel filtration Leishmania major

Specific Activity [micromol/min/mg]

Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
additional information
-
 relative activity, alpha-D-galacturonate 1-phosphate, 1.6%, forward reaction, + UTP, pH 7.8, 25°C Leishmania major
additional information
-
 relative activity, D-galactose 1-phosphate, 100%, forward reaction, + UTP, pH 7.8, 25°C Leishmania major
additional information
-
 relative activity, D-glucose 1-phosphate, 88%, forward reaction, + UTP, pH 7.8, 25°C Leishmania major
additional information
-
 relative activity, D-xylopyranose 1-phosphate, 12%, forward reaction, + UTP, pH 7.8, 25°C Leishmania major
additional information
-
 relative activity, UDP-alpha-D-galacturonate, 7.1%, reverse reaction, + diphosphate, pH 7.8, 25°C Leishmania major
additional information
-
 relative activity, UDP-beta-L-arabinopyranose, 33%, reverse reaction, + diphosphate, pH 7.8, 25°C Leishmania major
additional information
-
 relative activity, UDP-galactose, 83%, reverse reaction, + diphosphate, pH 7.8, 25°C Leishmania major
additional information
-
 relative activity, UDP-glucose, 100%, reverse reaction, + diphosphate, pH 7.8, 25°C Leishmania major

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
D-galactose 1-phosphate + UTP
-
 Leishmania major UDP-D-galactose + diphosphate
-
 ?
D-galacturonate 1-phosphate + UTP
-
 Leishmania major UDP-D-galacturonate + diphosphate
-
 ?
D-glucose 1-phosphate + UTP
-
 Leishmania major UDP-D-glucose + diphosphate
-
 ?
L-arabinopyranose 1-phosphate + UTP
-
 Leishmania major UDP-L-arabinopyranose + diphosphate
-
 ?
UTP + alpha-D-galacturonate
-
 Leishmania major diphosphate + UDP-alpha-D-galacturonate
-
 r
UTP + beta-L-arabinopyranose
-
 Leishmania major diphosphate + UDP-beta-L-arabinopyranose
-
 r
UTP + D-galactose 1-phosphate
-
 Leishmania major diphosphate + UDP-galactose
-
 r
UTP + D-glucose 1-phosphate
-
 Leishmania major diphosphate + UDP-glucose
-
 r
UTP + D-xylopyranose 1-phosphate
-
 Leishmania major diphosphate + UDP-D-xylopyranose
-
 r

Subunits

Subunits Comment Organism
monomer
-
 Leishmania major

Synonyms

Synonyms Comment Organism
UDP-sugar pyrophosphorylase
-
 Leishmania major
USP
-
 Leishmania major

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
25
-
 assay at Leishmania major
25
-
 activity assay Leishmania major

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
13
-
 UDP-D-galacturonate reverse reaction, pH 7.8, 25°C Leishmania major
13
-
 UDP-alpha-D-galacturonate reverse reaction, pH 7.8, 25°C Leishmania major
15
-
 diphosphate reverse reaction, UDP-alpha-D-galacturonate, pH 7.8, 25°C Leishmania major
15
-
 diphosphate reverse reaction, co-substrate: UDP-D-galacturonate, pH 7.8, 25°C Leishmania major
72
-
 UDP-beta-L-arabinopyranose reverse reaction, pH 7.8, 25°C Leishmania major
72
-
 UDP-L-arabinopyranose reverse reaction, pH 7.8, 25°C Leishmania major
81
-
 diphosphate reverse reaction, UDP-beta-L-arabinopyranose, pH 7.8, 25°C Leishmania major
81
-
 diphosphate reverse reaction, co-substrate: UDP-L-arabinopyranose, pH 7.8, 25°C Leishmania major
154
-
 UDP-galactose reverse reaction, pH 7.8, 25°C Leishmania major
154
-
 UDP-D-galactose reverse reaction, pH 7.8, 25°C Leishmania major
174
-
 diphosphate reverse reaction, UDP-galactose, pH 7.8, 25°C Leishmania major
174
-
 diphosphate reverse reaction, co-substrate: UDP-D-galactose, pH 7.8, 25°C Leishmania major
180
-
 UDP-glucose reverse reaction, pH 7.8, 25°C Leishmania major
180
-
 UDP-D-glucose reverse reaction, pH 7.8, 25°C Leishmania major
185
-
 diphosphate reverse reaction, UDP-glucose, pH 7.8, 25°C Leishmania major
185
-
 diphosphate reverse reaction, co-substrate: UDP-D-glucose, pH 7.8, 25°C Leishmania major
191
-
 D-glucose 1-phosphate forward reaction, pH 7.8, 25°C Leishmania major
201
-
 UTP forward reaction, D-galactose 1-phosphate, pH 7.8, 25°C Leishmania major
201
-
 UTP forward reaction, co-substrate: D-galactose 1-phosphate, pH 7.8, 25°C Leishmania major
209
-
 UTP forward reaction, D-glucose 1-phosphate, pH 7.8, 25°C Leishmania major
209
-
 UTP forward reaction, co-substrate: D-glucose 1-phosphate, pH 7.8, 25°C Leishmania major
219
-
 D-galactose 1-phosphate forward reaction, pH 7.8, 25°C Leishmania major

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
7.8
-
 assay at Leishmania major
7.8
-
 activity assay Leishmania major

General Information

General Information Comment Organism
physiological function USP is able to complement the growth defect of Escherichia coli galU mutant strain DEV6. This specific bacterial strain is unable to grow on agar-containing galactose as the only carbohydrate source due to a mutation in UDP-glucose pyrophosphorylase Leishmania major

kcat/KM [mM/s]

kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
16.7
-
 UDP-D-galacturonate reverse reaction, pH 7.8, 25°C Leishmania major
20.8
-
 diphosphate reverse reaction, co-substrate: UDP-D-galacturonate, pH 7.8, 25°C Leishmania major
81.2
-
 diphosphate reverse reaction, co-substrate: UDP-L-arabinopyranose, pH 7.8, 25°C Leishmania major
114
-
 D-glucose 1-phosphate forward reaction, pH 7.8, 25°C Leishmania major
198
-
 UDP-L-arabinopyranose reverse reaction, pH 7.8, 25°C Leishmania major
259
-
 D-galactose 1-phosphate forward reaction, pH 7.8, 25°C Leishmania major
492
-
 diphosphate reverse reaction, co-substrate: UDP-D-glucose, pH 7.8, 25°C Leishmania major
580
-
 diphosphate reverse reaction, co-substrate: UDP-D-galactose, pH 7.8, 25°C Leishmania major
1060
-
 UDP-D-glucose reverse reaction, pH 7.8, 25°C Leishmania major
1070
-
 UDP-D-galactose reverse reaction, pH 7.8, 25°C Leishmania major
1830
-
 UTP forward reaction, co-substrate: D-glucose 1-phosphate, pH 7.8, 25°C Leishmania major
2090
-
 UTP forward reaction, co-substrate: D-galactose 1-phosphate, pH 7.8, 25°C Leishmania major