Cloned (Comment) | Organism |
---|---|
The last 2.6 kbp of the human PIPkin IIbeta gene and the following 3.6 kbp are cloned and inserted into a plasmid which placed a FLAG-(His6)2 double-tag, followed by a puromycin selection cassette between them. Transfection of this into DT40 cells followed by puromycin selection produces cells which have the tag attached to the C-terminus of the protein when expressed by one of the two alleles. Experiments are conducted on one clonal line derived from this procedure calling JPR3. Successful integration of the tag at the correct genomic location was confirmed by PCR analysis. | Gallus gallus |
The last 2.6 kbp of the human PIPkin IIbeta gene and the following 3.6 kbp are cloned and inserted into a plasmid which placed a FLAG-(His6)2 double-tag, followed by a puromycin selection cassette between them. Transfection of this into DT40 cells followed by puromycin selection produces cells which have the tag attached to the C-terminus of the protein when expressed by one of the two alleles. Experiments are conducted on one clonal line derived from this procedure calling JPR3. Successful integration of the tag at the correct genomic location was confirmed by PCR analysis. | Homo sapiens |
Protein Variants | Comment | Organism |
---|---|---|
E300D | Site-directed mutagenesis is performed on the human Type IIbeta PIPkin cloned into pEGFP-C1 using the QuikChange mutagenesis method, to introduce an Glu300Asp amino acid substitution into the human nuclear localisation sequence producing the equivalent chicken sequence. | Gallus gallus |
E300D | Site-directed mutagenesis is performed on the human Type IIbeta PIPkin cloned into pEGFP-C1 using the QuikChange mutagenesis method, to introduce an Glu300Asp amino acid substitution into the human nuclear localisation sequence producing the equivalent chicken sequence. | Homo sapiens |
Localization | Comment | Organism | GeneOntology No. | Textmining |
---|---|---|---|---|
cytoplasm | small amount, it is unclear how much of PIPkin II in the cytoplasmic fraction can be accounted for by nuclear disintegration or by leakage of PIPkinIIbeta during the fractionation. | Homo sapiens | 5737 | - |
cytoplasm | small amount, it is unclear how much of PIPkin IIbeta in the cytoplasmic fraction can be accounted for by nuclear disintegration or by leakage of PIPkinIIbeta during the fractionation. | Gallus gallus | 5737 | - |
nucleus | - |
Gallus gallus | 5634 | - |
nucleus | - |
Homo sapiens | 5634 | - |
Molecular Weight [Da] | Molecular Weight Maximum [Da] | Comment | Organism |
---|---|---|---|
53000 | - |
FLAG-(His6)2 tagged PIPkin IIbeta, confirmed by western blotting | Gallus gallus |
53000 | - |
FLAG-(His6)2 tagged PIPkin IIbeta, confirmed by western blotting | Homo sapiens |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
ATP + 1-phosphatidyl-1D-myo-inositol 4-phosphate | Gallus gallus | 5' phosphorylation | ADP + 1-phosphatidyl-1D-myo-inositol 4,5-bisphosphate | - |
r | |
ATP + 1-phosphatidyl-1D-myo-inositol 4-phosphate | Homo sapiens | 5' phosphorylation | ADP + 1-phosphatidyl-1D-myo-inositol 4,5-bisphosphate | - |
r |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Gallus gallus | - |
Site-directed mutagenesis is performed on the human type IIbeta PIPkin cloned into pEGFP-C1 using QuikChange mutagenesis method, to introduce an Glu300Asp amino acid substitution into the human nuclear localisation sequence producing the equivalent chicken sequence 289-DRAEQEEMEVE(E300D)RAEDEE-306 | - |
Homo sapiens | P78356 | Site-directed mutagenesis is performed on the human type IIbeta PIPkin cloned into pEGFP-C1 using QuikChange mutagenesis method, to introduce an Glu300Asp amino acid substitution into the human nuclear localisation sequence producing the equivalent chicken sequence 289-DRAEQEEMEVE(E300D)RAEDEE-306 | - |
Purification (Comment) | Organism |
---|---|
affinity purification of FLAG-His6 tagged proteins using TALON beads | Gallus gallus |
affinity purification of FLAG-His6 tagged proteins using TALON beads | Homo sapiens |
Source Tissue | Comment | Organism | Textmining |
---|---|---|---|
cell culture | DT40 cell from chicken, that expresses active PIPkin IIbetA tagged at the C-terminus of Hef, a protein component of the Fanconi anemia-related tumor-suppressor complex. | Gallus gallus | - |
cell culture | DT40 cell from chicken, that expresses active PIPkin IIbetA tagged at the C-terminus of Hef, a protein component of the Fanconi anemia-related tumor-suppressor complex. | Homo sapiens | - |
Specific Activity Minimum [µmol/min/mg] | Specific Activity Maximum [µmol/min/mg] | Comment | Organism |
---|---|---|---|
additional information | - |
In three independent experiments, the ratio (tagged cells to wild-type) of immunoprecipitated enzyme activity, quantified by densitometry or by scintillation counting, is 2.37 +/-0.27 | Gallus gallus |
additional information | - |
In three independent experiments, the ratio (tagged cells to wild-type) of immunoprecipitated enzyme activity, quantified by densitometry or by scintillation counting, is 2.37 +/-0.27 | Homo sapiens |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
ATP + 1-phosphatidyl-1D-myo-inositol 4-phosphate | 5' phosphorylation | Gallus gallus | ADP + 1-phosphatidyl-1D-myo-inositol 4,5-bisphosphate | - |
r | |
ATP + 1-phosphatidyl-1D-myo-inositol 4-phosphate | 5' phosphorylation | Homo sapiens | ADP + 1-phosphatidyl-1D-myo-inositol 4,5-bisphosphate | - |
r |
Synonyms | Comment | Organism |
---|---|---|
PIPkinIIbeta | - |
Gallus gallus |
PIPkinIIbeta | - |
Homo sapiens |
type IIbeta phosphatidylinositol 5-phosphate 4-kinase | - |
Gallus gallus |
type IIbeta phosphatidylinositol 5-phosphate 4-kinase | - |
Homo sapiens |
type IIbeta PIPkin | - |
Gallus gallus |
type IIbeta PIPkin | - |
Homo sapiens |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
30 | - |
assay at | Gallus gallus |
30 | - |
assay at | Homo sapiens |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
7.4 | - |
assay at | Gallus gallus |
7.4 | - |
assay at | Homo sapiens |
Cofactor | Comment | Organism | Structure |
---|---|---|---|
ATP | - |
Gallus gallus | |
ATP | - |
Homo sapiens |