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Literature summary extracted from

  • Nurkanto, A.; Jeelani, G.; Yamamoto, T.; Hishiki, T.; Naito, Y.; Suematsu, M.; Hashimoto, T.; Nozaki, T.
    Biochemical, metabolomic, and genetic analyses of dephospho coenzyme A kinase involved in coenzyme A biosynthesis in the human enteric parasite Entamoeba histolytica (2018), Front. Microbiol., 9, 2902 .
    View publication on PubMedView publication on EuropePMC

Cloned(Commentary)

EC Number Cloned (Comment) Organism
2.7.1.24 gene DCPK1, phylogenetic analysis, recombinant His-tagged isozyme expression in Escherichia coli strain BL21(DE3), quantitative real-time PCR enzyme expression analysis, HA-, Myc-, or FLAG-tagged isozyme EhDPCK1 cannot be expressed Entamoeba histolytica
2.7.1.24 gene DCPK2, recombinant His-tagged isozyme expression in Escherichia coli strain BL21(DE3), quantitative real-time PCR enzyme expression analysis, recombinant expression of HA-tagged EhDPCK2 Entamoeba histolytica

Protein Variants

EC Number Protein Variants Comment Organism
2.7.1.24 additional information generation of trophozoites of Ehdpck1 and Ehdpck2 gene-silenced strains. Ehdpck1 and Ehdpck2 gene-silenced strains show significant growth defect in normal growth medium. The level of growth inhibition by Ehdpck2 gene silencing is more severe compared to that by Ehdpck1 gene silencing Entamoeba histolytica

Inhibitors

EC Number Inhibitors Comment Organism Structure
2.7.1.24 acetyl-CoA inhibition at higher concentrations; inhibition at higher concentrations Entamoeba histolytica
2.7.1.24 CoA inhibition at higher concentrations; inhibition at higher concentrations Entamoeba histolytica
2.7.1.24 malonyl-CoA inhibition at higher concentrations; inhibition at higher concentrations Entamoeba histolytica

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
2.7.1.24 additional information
-
 additional information both ATP and dephospho-CoA exhibit hyperbolic saturation kinetics when assayed over the range of 0.004-0.128 mM dephospho-CoA in the presence of 0.10 mM ATP and in the range of 0.001-0.10 mM ATP substrate in the presence of 0.128 mM dephospho-CoA Entamoeba histolytica
2.7.1.24 0.015
-
 ATP pH 8.0, 37°C, isozyme Ehdpck2 Entamoeba histolytica
2.7.1.24 0.0196
-
 ATP pH 8.0, 37°C, isozyme Ehdpck1 Entamoeba histolytica
2.7.1.24 0.0579
-
 3'-dephospho-CoA pH 8.0, 37°C, isozyme Ehdpck2 Entamoeba histolytica
2.7.1.24 0.114
-
 3'-dephospho-CoA pH 8.0, 37°C, isozyme Ehdpck1 Entamoeba histolytica

Localization

EC Number Localization Comment Organism GeneOntology No. Textmining
2.7.1.24 cytosol
-
 Entamoeba histolytica 5829
-
2.7.1.24 membrane
-
 Entamoeba histolytica 16020
-
2.7.1.24 additional information isozyme EhDPCK2 is located in both cytosol and organelles (or associated with the membrane) Entamoeba histolytica
-
-

Metals/Ions

EC Number Metals/Ions Comment Organism Structure
2.7.1.24 Ca2+ 1.0% activity compared to Mg2+ Entamoeba histolytica
2.7.1.24 Ca2+ 10.5% activity compared to Mg2+ Entamoeba histolytica
2.7.1.24 Co2+ 28.3% activity compared to Mg2+ Entamoeba histolytica
2.7.1.24 Co2+ 40.9% activity compared to Mg2+ Entamoeba histolytica
2.7.1.24 Cu2+ 53.2% activity compared to Mg2+ Entamoeba histolytica
2.7.1.24 Fe2+ 20.9% activity compared to Mg2+ Entamoeba histolytica
2.7.1.24 Fe2+ 61.5% activity compared to Mg2+ Entamoeba histolytica
2.7.1.24 Li+ 21.7% activity compared to Mg2+ Entamoeba histolytica
2.7.1.24 Li+ 4.9% activity compared to Mg2+ Entamoeba histolytica
2.7.1.24 Mg2+ dependent on Entamoeba histolytica
2.7.1.24 Mn2+ 43.8% activity compared to Mg2+ Entamoeba histolytica
2.7.1.24 Mn2+ 53.7% activity compared to Mg2+ Entamoeba histolytica
2.7.1.24 additional information EhDPCK1 and EhDPCK2 activities are Mg2+-dependent. Some cations (Zn2+ and Cu2+) can replace Mg2+ with relative activity higher than 50%. No activity is detected when Na+ or K+ are used instead of Mg2+ Entamoeba histolytica
2.7.1.24 Ni2+ 23.0% activity compared to Mg2+ Entamoeba histolytica
2.7.1.24 Ni2+ 45.2% activity compared to Mg2+ Entamoeba histolytica
2.7.1.24 Zn2+ 76.9% activity compared to Mg2+ Entamoeba histolytica
2.7.1.24 Zn2+ 78.3% activity compared to Mg2+ Entamoeba histolytica

Natural Substrates/ Products (Substrates)

EC Number Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
2.7.1.24 ATP + 3'-dephospho-CoA Entamoeba histolytica
-
 ADP + CoA
-
 ?

Organism

EC Number Organism UniProt Comment Textmining
2.7.1.24 Entamoeba histolytica C4LWC7 clonal strain HM-1:IMSS cl 6 and G3 strains
-
2.7.1.24 Entamoeba histolytica C4MB67 clonal strain HM-1:IMSS cl 6 and G3 strains
-

Purification (Commentary)

EC Number Purification (Comment) Organism
2.7.1.24 recombinant His-tagged isozyme from Escherichia coli strain BL21(DE3) by nickel affinity chromatography and dialysis Entamoeba histolytica

Source Tissue

EC Number Source Tissue Comment Organism Textmining
2.7.1.24 trophozoite
-
 Entamoeba histolytica
-

Specific Activity [micromol/min/mg]

EC Number Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
2.7.1.24 2.13
-
 purified recombinant isozyme Ehdpck1, pH 8.0, 37°C Entamoeba histolytica
2.7.1.24 2.54
-
 purified recombinant isozyme Ehdpck2, pH 8.0, 37°C Entamoeba histolytica

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
2.7.1.24 ATP + 3'-dephospho-CoA
-
 Entamoeba histolytica ADP + CoA
-
 ?
2.7.1.24 CTP + 3'-dephospho-CoA 15.4% activity compared to ATP Entamoeba histolytica CDP + CoA
-
 ?
2.7.1.24 CTP + 3'-dephospho-CoA 7.3% activity compared to ATP Entamoeba histolytica CDP + CoA
-
 ?
2.7.1.24 GTP + 3'-dephospho-CoA 32.6% activity compared to ATP Entamoeba histolytica GDP + CoA
-
 ?
2.7.1.24 GTP + 3'-dephospho-CoA 33% activity compared to ATP Entamoeba histolytica GDP + CoA
-
 ?
2.7.1.24 additional information pantothenate is no substrate Entamoeba histolytica ?
-
-
2.7.1.24 TTP + 3'-dephospho-CoA 24.1% activity compared to ATP Entamoeba histolytica TDP + CoA
-
 ?
2.7.1.24 TTP + 3'-dephospho-CoA 4.9% activity compared to ATP Entamoeba histolytica TDP + CoA
-
 ?
2.7.1.24 UTP + 3'-dephospho-CoA 2.6% activity compared to ATP Entamoeba histolytica UDP + CoA
-
 ?
2.7.1.24 UTP + 3'-dephospho-CoA 4.5% activity compared to ATP Entamoeba histolytica UDP + CoA
-
 ?

Subunits

EC Number Subunits Comment Organism
2.7.1.24 ? x * 23100, recombinant enzyme, SDS-PAGE, plus 2.6 kDa corresponding to the His-tag Entamoeba histolytica
2.7.1.24 ? x * 23900, recombinant enzyme, SDS-PAGE, plus 2.6 kDa corresponding to the His-tag Entamoeba histolytica

Synonyms

EC Number Synonyms Comment Organism
2.7.1.24 dephospho coenzyme A kinase
-
 Entamoeba histolytica
2.7.1.24 dephospho-CoA kinase
-
 Entamoeba histolytica
2.7.1.24 DPCK
-
 Entamoeba histolytica
2.7.1.24 Dpck1
-
 Entamoeba histolytica
2.7.1.24 Dpck2
-
 Entamoeba histolytica
2.7.1.24 EhDPCK1
-
 Entamoeba histolytica
2.7.1.24 EhDPCK2
-
 Entamoeba histolytica

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
2.7.1.24 37
-
 assay at Entamoeba histolytica

Turnover Number [1/s]

EC Number Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
2.7.1.24 0.96
-
 3'-dephospho-CoA pH 8.0, 37°C, isozyme Ehdpck2 Entamoeba histolytica
2.7.1.24 1.05
-
 ATP pH 8.0, 37°C, isozyme Ehddpck2 Entamoeba histolytica
2.7.1.24 1.41
-
 ATP pH 8.0, 37°C, isozyme Ehdpck1 Entamoeba histolytica
2.7.1.24 1.48
-
 3'-dephospho-CoA pH 8.0, 37°C, isozyme Ehdpck1 Entamoeba histolytica

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
2.7.1.24 8
-
 assay at Entamoeba histolytica

General Information

EC Number General Information Comment Organism
2.7.1.24 additional information intracellular metabolite extraction of wild-type expressing and silenced-mutant strain. DPCKs comprise three domains: the nucleotide-binding domain in parallel beta-sheet, the substrate-binding domain in alpha-helices, and the LID domain. All of these DPCK features are present in EhDPCK. Isozymes EhDPCK1 and 2 show striking similarity based on three-dimensional structure predicted by homology modeling, structural comparison of EhDPCK and its mammalian counterpart from mouse, overview Entamoeba histolytica

kcat/KM [mM/s]

EC Number kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
2.7.1.24 12.98
-
 3'-dephospho-CoA pH 8.0, 37°C, isozyme Ehdpck1 Entamoeba histolytica
2.7.1.24 16.58
-
 3'-dephospho-CoA pH 8.0, 37°C, isozyme Ehdpck2 Entamoeba histolytica
2.7.1.24 70
-
 ATP pH 8.0, 37°C, isozyme Ehdpck2 Entamoeba histolytica
2.7.1.24 71.43
-
 ATP pH 8.0, 37°C, isozyme Ehdpck1 Entamoeba histolytica