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Literature summary for 2.7.1.158 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)

Cloned (Comment) Organism
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)

Crystallization (Comment) Organism
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

Protein Variants Comment Organism
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

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

Molecular Weight [Da]

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

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
ATP + 1D-myo-inositol 1,3,4,5,6-pentakisphosphate Cryptococcus neoformans var. grubii
-
ADP + 1D-myo-inositol hexakisphosphate
-
?
ATP + 1D-myo-inositol 1,3,4,5,6-pentakisphosphate Cryptococcus neoformans var. grubii FGSC 9487
-
ADP + 1D-myo-inositol hexakisphosphate
-
?
ATP + 1D-myo-inositol 1,3,4,5,6-pentakisphosphate Cryptococcus neoformans var. grubii ATCC 208821
-
ADP + 1D-myo-inositol hexakisphosphate
-
?
ATP + 1D-myo-inositol 1,3,4,5,6-pentakisphosphate Cryptococcus neoformans var. grubii CBS 10515
-
ADP + 1D-myo-inositol hexakisphosphate
-
?

Organism

Organism UniProt Comment Textmining
Cryptococcus neoformans var. grubii J9VKS8 Filobasidiella neoformans var. grubii, serotype A
-
Cryptococcus neoformans var. grubii ATCC 208821 J9VKS8 Filobasidiella neoformans var. grubii, serotype A
-
Cryptococcus neoformans var. grubii CBS 10515 J9VKS8 Filobasidiella neoformans var. grubii, serotype A
-
Cryptococcus neoformans var. grubii FGSC 9487 J9VKS8 Filobasidiella neoformans var. grubii, serotype A
-

Purification (Commentary)

Purification (Comment) Organism
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)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
ATP + 1D-myo-inositol 1,3,4,5,6-pentakisphosphate
-
Cryptococcus neoformans var. grubii ADP + 1D-myo-inositol hexakisphosphate
-
?
ATP + 1D-myo-inositol 1,3,4,5,6-pentakisphosphate
-
Cryptococcus neoformans var. grubii FGSC 9487 ADP + 1D-myo-inositol hexakisphosphate
-
?
ATP + 1D-myo-inositol 1,3,4,5,6-pentakisphosphate
-
Cryptococcus neoformans var. grubii ATCC 208821 ADP + 1D-myo-inositol hexakisphosphate
-
?
ATP + 1D-myo-inositol 1,3,4,5,6-pentakisphosphate
-
Cryptococcus neoformans var. grubii CBS 10515 ADP + 1D-myo-inositol hexakisphosphate
-
?
additional information substrate binding structure, overview Cryptococcus neoformans var. grubii ?
-
-
additional information substrate binding structure, overview Cryptococcus neoformans var. grubii FGSC 9487 ?
-
-
additional information substrate binding structure, overview Cryptococcus neoformans var. grubii ATCC 208821 ?
-
-
additional information substrate binding structure, overview Cryptococcus neoformans var. grubii CBS 10515 ?
-
-

Subunits

Subunits Comment Organism
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
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

Synonyms Comment Organism
CnIpk1
-
Cryptococcus neoformans var. grubii
inositol 1,3,4,5,6-pentakisphosphate 2-kinase
-
Cryptococcus neoformans var. grubii
Ipk1
-
Cryptococcus neoformans var. grubii

Temperature Optimum [°C]

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

pH Optimum

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

Cofactor

Cofactor Comment Organism Structure
ATP
-
Cryptococcus neoformans var. grubii

General Information

General Information Comment Organism
additional information overall structure analysis of enzyme CnIpk1, conformational state and changes, overview Cryptococcus neoformans var. grubii