Cloned (Comment) | Organism |
---|---|
expression in Escherichia coli | Arabidopsis thaliana |
in Escherichia coli Rosetta, transformed with pET28a and pET28c containing 3 kinase cDNAs. Vector-encoded hexa-histidine (His) tags are added to all proteins. | Arabidopsis thaliana |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.042 | - |
1D-myo-inositol 1,3,4,6-tetrakisphosphate | - |
Arabidopsis thaliana |
Metals/Ions | Comment | Organism | Structure |
---|---|---|---|
LiCl | 10 mM in activity assay | Arabidopsis thaliana | |
MgCl2 | 6 mM in activity assay | Arabidopsis thaliana |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
1D-myo-inositol-1,3,4,5-tetrakisphosphate + ATP | Arabidopsis thaliana | enzyme displays isomerase activity | 1D-myo-inositol-1,3,4,6-tetrakisphosphate + ATP | - |
? | |
1D-myo-inositol-1,3,4-trisphosphate + ATP | Arabidopsis thaliana | - |
1D-myo-inositol-1,3,4,6-tetrakisphosphate + ADP | - |
? | |
1D-myo-inositol-1,4,6-trisphosphate + 1D-myo-inositol-3,4,6-trisphosphate + 2 ATP | Arabidopsis thaliana | racemic mixture | 2 1D-myo-inositol-1,3,4,6-tetrakisphosphate + 2 ADP | - |
? | |
1D-myo-inositol-3,4,5,6-tetrakisphosphate + ATP | Arabidopsis thaliana | enzyme displays isomerase activity | 1D-myo-inositol-1,3,4,6-tetrakisphosphate + ATP | - |
? | |
ATP + 1D-myo-inositol 1,3,4-trisphosphate | Arabidopsis thaliana | - |
ADP + 1D-myo-inositol 1,3,4,5-tetrakisphosphate | - |
? | |
ATP + 1D-myo-inositol 1,3,4-trisphosphate | Arabidopsis thaliana | - |
ADP + 1D-myo-inositol 1,3,4,6-tetrakisphosphate | - |
? | |
ATP + 1D-myo-inositol 1,3,4-trisphosphate | Arabidopsis thaliana | The substrate is converted to InsP4 but not to InsP5 | ADP + 1D-myo-inositol 1,3,4,6-tetrakisphosphate | - |
? | |
ATP + 1D-myo-inositol 1,4,6-trisphosphate + 1D-myo-inositol 3,4,6-trisphosphate | Arabidopsis thaliana | racemic mix of phosphates | ADP + inositol tetrakisphosphate | - |
? | |
ATP + 1D-myo-inositol 3,4,5,6-tetrakisphosphate | Arabidopsis thaliana | - |
ADP + 1D-myo-inositol 1,3,4,5,6-pentakisphosphate | - |
? | |
additional information | Arabidopsis thaliana | AtITPK4 differs from other family members in that it not display inositol 3,4,5,6-tetrakisphosphate 1-kinase activity. It displays inositol 1,4,5,6-tetrakisphosphate and inositol-1,3,4,5-tetrakisphosphate isomerase activity. With inositol tetrakisphosphate substrates, ITPK4 displays an isomerase activity (in the absence of inositol pentakisphosphate product) interpreting as phosphatase and kinase activity. AtITPK4 shows isomerase activity against other substrates. Removal of the 1- or 3-phosphate of inositol-1,3,4,6-tetrakisphosphate and subsequent rephosphorylation at the same position generates inositol-1,3,4,6-tetrakisphosphate. | ? | - |
? | |
additional information | Arabidopsis thaliana | enzyme displays both kinase and phosphatase activity. | ? | - |
? |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Arabidopsis thaliana | O80568 | - |
- |
Arabidopsis thaliana | O80568 | AtITPK4 protein is distinguished from the other proteins by an extended N-terminal sequence (39 residues longer than that of AtITPK2). A blast search with amino acids 1-80 of AtITPK4 as query fails to identify significant homology to any proteins of other known functional annotation. In alignment, AtITPK4 has an extra 121 amino acids N-terminal to the start of the Entamoeba enzyme. AtITPK4 shows 14.9% identity and 25.7% similarity to the Entamoeba enzyme in a pairwise alignment. | - |
Arabidopsis thaliana | O81893 | enzyme belongs to a larger family of ATP-grasp fold proteins | - |
Arabidopsis thaliana | Q9SBA5 | enzyme belongs to a larger family of ATP-grasp fold proteins | - |
Arabidopsis thaliana | Q9SUG3 | enzyme belongs to a larger family of ATP-grasp fold proteins | - |
Purification (Comment) | Organism |
---|---|
Cells are pelleted and resuspended in buffer, sonicated, centrifuged and the supernatant of the lysate is retained. Ni2+-nitrilotriacetate resin prewashed in NiSO4 is added to the lysate and incubated. The resin is washed twice with 10 ml of 50 mM NaH2PO4, 300 mM NaCl and 20 mM imidazole (pH 8) to elute non-specifically bound proteins, and washed with of NaH2PO4, 300 mM NaCl and 250 mM imidazole to elute AtIPK1 protein. Bacterial lysates and protein fractions from the purification are analysed by SDS-PAGE. Protein concentration is determined by the Bradford assay. | Arabidopsis thaliana |
Cells are pelleted and resuspended in buffer, sonicated, centrifuged and the supernatant of the lysate is retained. Ni2+-nitrilotriacetate resin prewashed in NiSO4 is added to the lysate and incubated. The resin is washed twice with 10 ml of 50 mM NaH2PO4, 300 mM NaCl and 20 mM imidazole (pH 8) to elute non-specifically bound proteins, and washed with of NaH2PO4, 300 mM NaCl and 250 mM imidazole to elute AtIPK1 protein. Bacterial lysates and protein fractions from the purification are analysed by SDS-PAGE. Protein concentration is determined by the Bradford method. | Arabidopsis thaliana |
Cells are pelleted and resuspended in buffer, sonicated, centrifuged and the supernatant of the lysate is retained. Ni2+-nitrilotriacetate resin prewashed in NiSO4 is added to the lysate and incubated. The resin is washed twice with 10 ml of 50 mM NaH2PO4, 300 mM NaCl and 20 mM imidazole (pH 8) to elute non-specifically bound proteins, and washed with of NaH2PO4, 300 mM NaCl and 250 mM imidazole to elute AtIPK1 protein. Bacterial lysates and protein fractions from the purification are analysed by SDS/PAGE. Protein concentration is determined by Bradford assay. | Arabidopsis thaliana |
Cells are pelleted and resuspended in buffer, sonicated, centrifuged and the supernatant of the lysate is retained. Ni2+-nitrilotriacetate resin prewashed in NiSO4 is added to the lysate and incubated. The resin is washed twice with 10 ml of 50 mM NaH2PO4, 300 mM NaCl and 20 mM imidazole (pH 8) to elute non-specifically bound proteins, and washed with of NaH2PO4, 300 mM NaCl and 250 mM imidazole to elute AtIPK1 protein. Bacterial lysates and protein fractions from the purification are analysed by SDS/PAGE. Protein concentration is determined by the Bradford method. | Arabidopsis thaliana |
Source Tissue | Comment | Organism | Textmining |
---|---|---|---|
anther | - |
Arabidopsis thaliana | - |
carpel | - |
Arabidopsis thaliana | - |
cauline leaf | - |
Arabidopsis thaliana | - |
cauline leaf | weak expression | Arabidopsis thaliana | - |
flower | - |
Arabidopsis thaliana | - |
flower | expressed in male and female organs of young and mature flowers | Arabidopsis thaliana | - |
flower bud | young, AtITPK4 transcript is expressed throughout early flower development which culminates in the opening of the flower bud | Arabidopsis thaliana | - |
leaf | - |
Arabidopsis thaliana | - |
leaf | weak expression, vasculature | Arabidopsis thaliana | - |
petal | - |
Arabidopsis thaliana | - |
pollen | - |
Arabidopsis thaliana | - |
root | - |
Arabidopsis thaliana | - |
sepal | - |
Arabidopsis thaliana | - |
silique | - |
Arabidopsis thaliana | - |
silique | weak expression, revealed expression in the seed and pod wall | Arabidopsis thaliana | - |
stamen | - |
Arabidopsis thaliana | - |
stem | - |
Arabidopsis thaliana | - |
Specific Activity Minimum [µmol/min/mg] | Specific Activity Maximum [µmol/min/mg] | Comment | Organism |
---|---|---|---|
additional information | - |
not detected in mature flowers, but expression of trancript is confirmed by mRNA in situ analysis | Arabidopsis thaliana |
0.0044 | - |
- |
Arabidopsis thaliana |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
1D-myo-inositol-1,3,4,5-tetrakisphosphate + ATP | enzyme displays isomerase activity | Arabidopsis thaliana | 1D-myo-inositol-1,3,4,6-tetrakisphosphate + ATP | - |
? | |
1D-myo-inositol-1,3,4-trisphosphate + ATP | - |
Arabidopsis thaliana | 1D-myo-inositol-1,3,4,6-tetrakisphosphate + ADP | - |
? | |
1D-myo-inositol-1,4,5,6-tetrakisphosphate + ATP | AtITPK4 is more active against 1D-myo-inositol-1,4,5,6-tetrakisphosphate than against 1D-myo-inositol-3,4,5,6-tetrakisphosphate. | Arabidopsis thaliana | 1D-myo-inositol-1,3,4,6-tetrakisphosphate + ATP | - |
? | |
1D-myo-inositol-1,4,6-trisphosphate + 1D-myo-inositol-3,4,6-trisphosphate + 2 ATP | racemic mixture | Arabidopsis thaliana | 2 1D-myo-inositol-1,3,4,6-tetrakisphosphate + 2 ADP | - |
? | |
1D-myo-inositol-3,4,5,6-tetrakisphosphate + ATP | enzyme displays isomerase activity | Arabidopsis thaliana | 1D-myo-inositol-1,3,4,6-tetrakisphosphate + ATP | - |
? | |
ATP + 1D-myo-inositol 1,3,4-trisphosphate | - |
Arabidopsis thaliana | ADP + 1D-myo-inositol 1,3,4,5-tetrakisphosphate | - |
? | |
ATP + 1D-myo-inositol 1,3,4-trisphosphate | - |
Arabidopsis thaliana | ADP + 1D-myo-inositol 1,3,4,6-tetrakisphosphate | - |
? | |
ATP + 1D-myo-inositol 1,3,4-trisphosphate | The substrate is converted to InsP4 but not to InsP5 | Arabidopsis thaliana | ADP + 1D-myo-inositol 1,3,4,6-tetrakisphosphate | - |
? | |
ATP + 1D-myo-inositol 1,4,6-trisphosphate + 1D-myo-inositol 3,4,6-trisphosphate | racemic mix of phosphates | Arabidopsis thaliana | ADP + inositol tetrakisphosphate | - |
? | |
ATP + 1D-myo-inositol 3,4,5,6-tetrakisphosphate | - |
Arabidopsis thaliana | ADP + 1D-myo-inositol 1,3,4,5,6-pentakisphosphate | - |
? | |
ATP + 1D-myo-inositol-1,3,4-trisphosphate | - |
Arabidopsis thaliana | ? | - |
? | |
additional information | the enzyme displays inositol 1,4,5,6-tetrakisphosphate and inositol 1,3,4,5-tetrakisphosphate isomerase activity. It lacks the Ins(3,4,5,6)P4 1-kinase | Arabidopsis thaliana | ? | - |
? | |
additional information | AtITPK4 differs from other family members in that it not display inositol 3,4,5,6-tetrakisphosphate 1-kinase activity. It displays inositol 1,4,5,6-tetrakisphosphate and inositol-1,3,4,5-tetrakisphosphate isomerase activity. With inositol tetrakisphosphate substrates, ITPK4 displays an isomerase activity (in the absence of inositol pentakisphosphate product) interpreting as phosphatase and kinase activity. AtITPK4 shows isomerase activity against other substrates. Removal of the 1- or 3-phosphate of inositol-1,3,4,6-tetrakisphosphate and subsequent rephosphorylation at the same position generates inositol-1,3,4,6-tetrakisphosphate. | Arabidopsis thaliana | ? | - |
? | |
additional information | enzyme displays both kinase and phosphatase activity. | Arabidopsis thaliana | ? | - |
? |
Synonyms | Comment | Organism |
---|---|---|
At2G43980 | - |
Arabidopsis thaliana |
AtITPK1 | - |
Arabidopsis thaliana |
AtITPK2 | - |
Arabidopsis thaliana |
AtITPK3 | - |
Arabidopsis thaliana |
AtITPK4 | - |
Arabidopsis thaliana |
inositol 1,3,4-trisphosphate 5/6-kinase | - |
Arabidopsis thaliana |
inositol 1,3,4-trisphosphate 5/6-kinase 4 | - |
Arabidopsis thaliana |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
30 | - |
assay at | Arabidopsis thaliana |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
7.5 | - |
assay at | Arabidopsis thaliana |
Cofactor | Comment | Organism | Structure |
---|---|---|---|
ATP | - |
Arabidopsis thaliana | |
ATP | commentary | Arabidopsis thaliana |