EC Number | Cloned (Comment) | Organism |
---|---|---|
2.7.1.158 | gene IPK1, recombinant expression of N-terminally His5-tagged wild-type and mutant enzymes in Escherichia coli strain BL21(DE3) | Cryptococcus neoformans var. grubii |
EC Number | Crystallization (Comment) | Organism |
---|---|---|
2.7.1.158 | purified recombinant detagged wild-type and mutant enzymes in complex with IP6 and ADP, sitting drop vapor diffusion method, mixing of 20 mg/ml protein solution with crystallization solution containing 0.1 M MES, pH 6.0, 2.1-2.3 M ammonium sulfate, and 10-50 mM TCEP, 22°C, 20% v/v ethylene glycol is used as a cryoprotectant, X-ray diffraction structure determination and analysis at 2.35-2.5 A resolution | Cryptococcus neoformans var. grubii |
EC Number | Protein Variants | Comment | Organism |
---|---|---|---|
2.7.1.158 | Q64A/E66A/E67A | site-directed mutagenesis, mutant structure analysis with bound reaction products IP6 and ADP. CnIpk1 mutant Q64A/E66A/E67A forms a crystal but only in the presence of the products IP6 and ADP | Cryptococcus neoformans var. grubii |
EC Number | Metals/Ions | Comment | Organism | Structure |
---|---|---|---|---|
2.7.1.158 | Mg2+ | required, binding structure, overview | Cryptococcus neoformans var. grubii |
EC Number | Molecular Weight [Da] | Molecular Weight Maximum [Da] | Comment | Organism |
---|---|---|---|---|
2.7.1.158 | 41000 | - |
gel filtration | Cryptococcus neoformans var. grubii |
EC Number | Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|---|
2.7.1.158 | ATP + 1D-myo-inositol 1,3,4,5,6-pentakisphosphate | Cryptococcus neoformans var. grubii | - |
ADP + 1D-myo-inositol hexakisphosphate | - |
? | |
2.7.1.158 | ATP + 1D-myo-inositol 1,3,4,5,6-pentakisphosphate | Cryptococcus neoformans var. grubii FGSC 9487 | - |
ADP + 1D-myo-inositol hexakisphosphate | - |
? | |
2.7.1.158 | ATP + 1D-myo-inositol 1,3,4,5,6-pentakisphosphate | Cryptococcus neoformans var. grubii ATCC 208821 | - |
ADP + 1D-myo-inositol hexakisphosphate | - |
? | |
2.7.1.158 | ATP + 1D-myo-inositol 1,3,4,5,6-pentakisphosphate | Cryptococcus neoformans var. grubii CBS 10515 | - |
ADP + 1D-myo-inositol hexakisphosphate | - |
? |
EC Number | Organism | UniProt | Comment | Textmining |
---|---|---|---|---|
2.7.1.158 | Cryptococcus neoformans var. grubii | J9VKS8 | Filobasidiella neoformans var. grubii, serotype A | - |
2.7.1.158 | Cryptococcus neoformans var. grubii ATCC 208821 | J9VKS8 | Filobasidiella neoformans var. grubii, serotype A | - |
2.7.1.158 | Cryptococcus neoformans var. grubii CBS 10515 | J9VKS8 | Filobasidiella neoformans var. grubii, serotype A | - |
2.7.1.158 | Cryptococcus neoformans var. grubii FGSC 9487 | J9VKS8 | Filobasidiella neoformans var. grubii, serotype A | - |
EC Number | Purification (Comment) | Organism |
---|---|---|
2.7.1.158 | recombinant N-terminally His5-tagged wild-type and mutant enzymes from Escherichia coli strain BL21(DE3) by nickel affinity chromatography, dialysis, and tag cleavage through TEV protease, followed by another step of nickel affinity chromatography, and gel filtration | Cryptococcus neoformans var. grubii |
EC Number | Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|---|
2.7.1.158 | ATP + 1D-myo-inositol 1,3,4,5,6-pentakisphosphate | - |
Cryptococcus neoformans var. grubii | ADP + 1D-myo-inositol hexakisphosphate | - |
? | |
2.7.1.158 | ATP + 1D-myo-inositol 1,3,4,5,6-pentakisphosphate | - |
Cryptococcus neoformans var. grubii FGSC 9487 | ADP + 1D-myo-inositol hexakisphosphate | - |
? | |
2.7.1.158 | ATP + 1D-myo-inositol 1,3,4,5,6-pentakisphosphate | - |
Cryptococcus neoformans var. grubii ATCC 208821 | ADP + 1D-myo-inositol hexakisphosphate | - |
? | |
2.7.1.158 | ATP + 1D-myo-inositol 1,3,4,5,6-pentakisphosphate | - |
Cryptococcus neoformans var. grubii CBS 10515 | ADP + 1D-myo-inositol hexakisphosphate | - |
? | |
2.7.1.158 | additional information | substrate binding structure, overview | Cryptococcus neoformans var. grubii | ? | - |
- |
|
2.7.1.158 | additional information | substrate binding structure, overview | Cryptococcus neoformans var. grubii FGSC 9487 | ? | - |
- |
|
2.7.1.158 | additional information | substrate binding structure, overview | Cryptococcus neoformans var. grubii ATCC 208821 | ? | - |
- |
|
2.7.1.158 | additional information | substrate binding structure, overview | Cryptococcus neoformans var. grubii CBS 10515 | ? | - |
- |
EC Number | Subunits | Comment | Organism |
---|---|---|---|
2.7.1.158 | monomer | 1 * 41000, there are four monomers in the asymmetric unit, but size-exclusion chromatographic analysis indicates that an estimated molecular weight of CnIpk1 corresponds to 41 kDa, which is equivalent to a monomer. The four monomers in the asymmetric unit do not form any noticeable oligomeric conformation in a symmetric manner, which indicated that CnIpk1 is a monomeric enzyme | Cryptococcus neoformans var. grubii |
2.7.1.158 | More | enzyme CnIpk1 exhibits two structural lobes: the N-terminal lobe (N-lobe, residues Met1-Glu134) and the C-terminal lobe (C-lobe, residues Ile146-Arg415) with an 11-residue loop between the lobes. Specifically, the N-lobe forms an alpha/beta fold with five antiparallel beta-strands in an 1-2-3-5-4 order and surrounding alpha-helices, structure overview | Cryptococcus neoformans var. grubii |
EC Number | Synonyms | Comment | Organism |
---|---|---|---|
2.7.1.158 | CnIpk1 | - |
Cryptococcus neoformans var. grubii |
2.7.1.158 | inositol 1,3,4,5,6-pentakisphosphate 2-kinase | - |
Cryptococcus neoformans var. grubii |
2.7.1.158 | Ipk1 | - |
Cryptococcus neoformans var. grubii |
EC Number | Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|---|
2.7.1.158 | 22 | - |
assay at | Cryptococcus neoformans var. grubii |
EC Number | pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|---|
2.7.1.158 | 7.5 | - |
assay at | Cryptococcus neoformans var. grubii |
EC Number | Cofactor | Comment | Organism | Structure |
---|---|---|---|---|
2.7.1.158 | ATP | - |
Cryptococcus neoformans var. grubii |
EC Number | General Information | Comment | Organism |
---|---|---|---|
2.7.1.158 | additional information | overall structure analysis of enzyme CnIpk1, conformational state and changes, overview | Cryptococcus neoformans var. grubii |