Cloned (Comment) | Organism |
---|---|
gene PH0925, recombinant expression of His-tagged enzyme PH0925 in Escherichia coli strain BL21-Codon Plus(DE3)-RIL | Pyrococcus horikoshii |
Protein Variants | Comment | Organism |
---|---|---|
additional information | construction of several truncated mutants of the PH0925 protein, that show altered kinetics compared to the wild-type enzyme, overview | Pyrococcus horikoshii |
Inhibitors | Comment | Organism | Structure |
---|---|---|---|
EDTA | complete inhibition | Pyrococcus horikoshii | |
additional information | the C-terminal 114 residue region of the PH0925 protein inhibits the Man-1-P GTase activity. The phosphomannose isomerase activity is abolished by deletion of the C-terminal 14 residues | Pyrococcus horikoshii |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.0026 | - |
GTP | pH 7.6, 85°C, recombinant enzyme | Pyrococcus horikoshii | |
0.0161 | - |
alpha-D-mannose 1-phosphate | pH 7.6, 85°C, recombinant enzyme | Pyrococcus horikoshii |
Metals/Ions | Comment | Organism | Structure |
---|---|---|---|
Ca2+ | activates | Pyrococcus horikoshii | |
Co2+ | activates | Pyrococcus horikoshii | |
Cu2+ | best activating metal ion | Pyrococcus horikoshii | |
Mg2+ | highly activates sugar-1-P NTase activity with substrate Man-1-P, obly slightly activating with substrates Glc-1-P and GlcN-1-P | Pyrococcus horikoshii | |
Mn2+ | activates | Pyrococcus horikoshii | |
additional information | the metal cofactor requirements of the multifunctional enzyme differ with the substrates used, overview | Pyrococcus horikoshii | |
Ni2+ | activates slightly | Pyrococcus horikoshii | |
Zn2+ | highly activating | Pyrococcus horikoshii |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
GTP + alpha-D-mannose 1-phosphate | Pyrococcus horikoshii | - |
diphosphate + GDP-mannose | - |
? | |
additional information | Pyrococcus horikoshii | a bifunctional enzyme with phosphomannose isomerase (PMI), EC 5.3.1.8, and mannose-1-phosphate guanylyltransferase (Man-1-P GTase), EC 2.7.7.13, activities, which can synthesize GDP-mannose when accompanied by a phosphomannomutase/phosphoglucomutase (PMM/PGM) enzyme (PH0923). PH0925 protein is a thermostable enzyme with both PMI and multiple sugar-1-P NTase, cf. EC 2.7.7.37, activities. Substrate specificity of the sugar-1-P NTase activity of the PH0925 protein, overview | ? | - |
? |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Pyrococcus horikoshii | O58649 | - |
- |
Pyrococcus horikoshii ATCC 700860 | O58649 | - |
- |
Purification (Comment) | Organism |
---|---|
recombinant His-tagged PH0925 protein from Escherichia coli strain BL21-Codon Plus(DE3)-RIL by nickel affinity chromatography | Pyrococcus horikoshii |
Specific Activity Minimum [µmol/min/mg] | Specific Activity Maximum [µmol/min/mg] | Comment | Organism |
---|---|---|---|
10.2 | - |
with GTP and alpha-D-mannose 1-phosphate, purified His-tagged recombinant enzyme, pH 7.6, 85°C | Pyrococcus horikoshii |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
GTP + alpha-D-mannose 1-phosphate | - |
Pyrococcus horikoshii | diphosphate + GDP-mannose | - |
? | |
additional information | a bifunctional enzyme with phosphomannose isomerase (PMI), EC 5.3.1.8, and mannose-1-phosphate guanylyltransferase (Man-1-P GTase), EC 2.7.7.13, activities, which can synthesize GDP-mannose when accompanied by a phosphomannomutase/phosphoglucomutase (PMM/PGM) enzyme (PH0923). PH0925 protein is a thermostable enzyme with both PMI and multiple sugar-1-P NTase, cf. EC 2.7.7.37, activities. Substrate specificity of the sugar-1-P NTase activity of the PH0925 protein, overview | Pyrococcus horikoshii | ? | - |
? | |
additional information | no activity with dATP, dCTP, and dTTP, and no activity with fructose-1-phosphate, alpha-D-galactose-1-phosphate, and N-acetyl-D-glucosamine-1-phosphate | Pyrococcus horikoshii | ? | - |
? |
Synonyms | Comment | Organism |
---|---|---|
More | cf. EC 2.7.7.13 and EC 2.7.7.34 | Pyrococcus horikoshii |
multiple sugar-1-P NTase | - |
Pyrococcus horikoshii |
multiple sugar-1-phosphate nucleotidylyltransferase | - |
Pyrococcus horikoshii |
PH0925 | - |
Pyrococcus horikoshii |
PH0925 protein | - |
Pyrococcus horikoshii |
sugar-1-phosphate nucleotidylyltransferase | - |
Pyrococcus horikoshii |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
95 | - |
- |
Pyrococcus horikoshii |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
7.6 | - |
assay at | Pyrococcus horikoshii |
General Information | Comment | Organism |
---|---|---|
evolution | the enzyme belongs to the mannose-6-phosphate isomerase type 2 family | Pyrococcus horikoshii |
additional information | the sugar-1-phosphate nucleotidylyltransferase activity is located in the region from the N-terminus to the 345th residue, the Man-1-P GTase activity is located in the C-terminal 114 residue region of the PH0925 protein, the phosphomannose isomerase requires the the C-terminal 14 residues | Pyrococcus horikoshii |