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2.7.1.159: inositol-1,3,4-trisphosphate 5/6-kinase

This is an abbreviated version!
For detailed information about inositol-1,3,4-trisphosphate 5/6-kinase, go to the full flat file.

Word Map on EC 2.7.1.159

Reaction

ATP
+
1D-myo-inositol 1,3,4-trisphosphate
=
ADP
 +
1D-myo-inositol 1,3,4,5-tetrakisphosphate

Synonyms

1,3,4-trisphosphate 5/6-kinase, 5/6 kinase, 5/6-kinase, At2G43980, AtITPK1, AtITPK2, AtITPK3, AtITPK4, GmItpk2, inositol 1,3,4 tris phosphate 5/6 kinase-2, inositol 1,3,4 trisphosphate kinase-2, inositol 1,3,4-triphosphate 5/6 kinase, inositol 1,3,4-triphosphate 5/6-kinase, inositol 1,3,4-trisphosphate 5/6 kinase-2, inositol 1,3,4-trisphosphate 5/6-kinase, inositol 1,3,4-trisphosphate 5/6-kinase 1, inositol 1,3,4-trisphosphate 5/6-kinase 4, inositol 1,3,4-trisphosphate 5/6-kinase6, inositol 1,4,5 trisphosphate 3/6 kinase, inositol-1,3,4-trisphosphate kinase, Ins(1,3,4)P3 5/6-kinase, IP56K, Ipk2, ITPK, Itpk-1, ITPK1, ITPK2, ITPK3, More, Os09g0518700, OsITL1, OsITPK6

ECTree

     2 Transferases
         2.7 Transferring phosphorus-containing groups
             2.7.1 Phosphotransferases with an alcohol group as acceptor
                2.7.1.159 inositol-1,3,4-trisphosphate 5/6-kinase

Purification

Purification on EC 2.7.1.159 - inositol-1,3,4-trisphosphate 5/6-kinase

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PURIFICATION (Commentary)
ORGANISM
UNIPROT
LITERATURE
12900fold from brain by ion exchange chromatography, and heparin/inositol hexakisphosphate affinity chromatography
-
26171fold from liver by polyethylene glycol precipitation, ion exchange chromatography and heparin and inositol hexakisphosphate affinity chromatograpies
-
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.
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.
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.
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.
partially as COP9 signalosome complex and further as a single protein
-
recombinant enzyme from insect Sf21 cells to homogeneity
-
recombinant enzyme from insect Sf21 cells, recombinant enzyme partially from 293 cells by ion exchange chromatography
-
recombinant GST-tagged enzyme from Escherichia coli by glutathione affinity chromatography and gel filtration
recombinant GST-tagged enzyme, the GST-tag is cleaved off by treatment with GST-3Cpro
recombinant His-tagged enzyme from Escherichia coli by adsorption and calmodulin affinity chromatography
recombinant His-tagged enzyme from Escherichia coli strain BL21
recombinant His5-tagged GmITPK2 from Escherichia coli strain BL21(DE3) by nickel affinity chromatography