Cloned (Comment) | Organism |
---|---|
expression of mutant cDNA fragment encoding for residues 185-459 of the catalytic domain of isozyme A as unlabeled or selenomethionine-labeled GST fusion proteins in Escherichia coli | Rattus norvegicus |
Crystallization (Comment) | Organism |
---|---|
purified recombinant unlabeled or selennomethionine-labeled mutant catalytic domain residues 185-459, 10 mg/ml protein in 25 mM Tris-HCl, pH 8.0, 150 mM NaCl, and 10 mM DTT, crystallization solution contains 2.2 M ammonium sulfate, 0.2 M sodium formate, 10 mM DTT, 10 mM ADP, 10 mM 1D-myo-inositol 1,3,4,5-tetrakisphosphate, and 10 mM MgCl2, cryoprotection of crystals by successive transfer into 5-15% glycerol in mother liquor, X-ray diffraction structure determination and analysis at 2.2 A resolution | Rattus norvegicus |
Protein Variants | Comment | Organism |
---|---|---|
D423N | crystal structure determination and analysis | Rattus norvegicus |
L217M | crystal structure determination and analysis | Rattus norvegicus |
Metals/Ions | Comment | Organism | Structure |
---|---|---|---|
Mg2+ | - |
Rattus norvegicus |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Rattus norvegicus | - |
isozyme A | - |
Purification (Comment) | Organism |
---|---|
unlabeled or selenomethionine-labeled recombinant residues 185-459 of the catalytic domain of isozyme A fused to GST from Escherichia coli by glutathione affinity chromatography, removal of the GST-tag by recombinant TEV protease, further purification by a second step of glutathione affinity chromatography, adsorption chromatography, and gel filtration | Rattus norvegicus |
Reaction | Comment | Organism | Reaction ID |
---|---|---|---|
ATP + 1D-myo-inositol 1,4,5-trisphosphate = ADP + 1D-myo-inositol 1,3,4,5-tetrakisphosphate | catalytic core structure, substrate binding and catalytic mechanism, structure-function realtionship | Rattus norvegicus |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
ATP + 1D-myo-inositol 1,4,5-trisphosphate | - |
Rattus norvegicus | ADP + 1D-myo-inositol 1,3,4,5-tetrakisphosphate | - |
? | |
additional information | the enzyme contains a catalytic domain and a 4-helix substrate binding domain, both are in an open conformation with respect to each other, thus substrate recognition and catalysis involve a dynamic conformational cycle | Rattus norvegicus | ? | - |
? |
Subunits | Comment | Organism |
---|---|---|
More | the enzyme contains a catalytic domain and a 4-helix substrate binding domain, both are in an open conformation with respect to each other, thus substrate recognition and catalysis involve a dynamic conformational cycle | Rattus norvegicus |
Synonyms | Comment | Organism |
---|---|---|
inositol 1,4,5-trisphosphate 3-kinase | - |
Rattus norvegicus |
IP3KA | - |
Rattus norvegicus |
More | the enzyme is a member of the protein kinase superfamily | Rattus norvegicus |
Cofactor | Comment | Organism | Structure |
---|---|---|---|
ATP | binding site structure | Rattus norvegicus |