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Literature summary extracted from

  • Oh, J.; Lee, D.G.; Bahn, Y.S.; Rhee, S.
    Crystal structure of inositol 1,3,4,5,6-pentakisphosphate 2-kinase from Cryptococcus neoformans (2017), J. Struct. Biol., 200, 118-123 .
    View publication on PubMed

Cloned(Commentary)

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

Crystallization (Commentary)

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

Protein Variants

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

Metals/Ions

EC Number Metals/Ions Comment Organism Structure
2.7.1.158 Mg2+ required, binding structure, overview Cryptococcus neoformans var. grubii

Molecular Weight [Da]

EC Number Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
2.7.1.158 41000
-
 gel filtration Cryptococcus neoformans var. grubii

Natural Substrates/ Products (Substrates)

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
-
 ?

Organism

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
-

Purification (Commentary)

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

Substrates and Products (Substrate)

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 ?
-
-

Subunits

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

Synonyms

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

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
2.7.1.158 22
-
 assay at Cryptococcus neoformans var. grubii

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
2.7.1.158 7.5
-
 assay at Cryptococcus neoformans var. grubii

Cofactor

EC Number Cofactor Comment Organism Structure
2.7.1.158 ATP
-
 Cryptococcus neoformans var. grubii

General Information

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