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Literature summary extracted from

  • Machado, A.; Silva, M.; Iulek, J.
    Expression, purification, enzymatic characterization and crystallization of glyceraldehyde-3-phosphate dehydrogenase from Naegleria gruberi, the first one from phylum Percolozoa (2016), Protein Expr. Purif., 127, 125-130 .
    View publication on PubMed

Activating Compound

EC Number Activating Compound Comment Organism Structure
1.2.1.12 arsenate best at 10 mM and up to 30 mM Naegleria gruberi

Cloned(Commentary)

EC Number Cloned (Comment) Organism
1.2.1.12 GAPDH gene, DNA and amino acid sequence determination and analysis, phylogenetic analysis, recombinant expression of N-terminally His-tagged enzyme in Escherichia coli strain BL21(DE3) Naegleria gruberi
1.2.1.107 expressed in Escherichia coli BL21(DE3) cells Naegleria gruberi

Crystallization (Commentary)

EC Number Crystallization (Comment) Organism
1.2.1.12 purified recombinant N-terminally His-tagged enzyme, hanging drop vapor diffusion method, mixing of 10 mg/ml protein with reservoir solution containing either 12.9% w/v PEG 2000, 50 mM acetate buffer, pH 5.2, and 6.4% w/v PEG 200, or 2.5% w/v PEG 1000, 50 mM acetate buffer, pH 4.8, and 25.7% w/v PEG 2000 MME, 18°C, 3 days, X-ray diffraction structure determination and analysis at 2.40 A and 1.94 A resolutions, respectively Naegleria gruberi
1.2.1.107 hanging drop vapor diffusion method, using either 12.9% (w/w) PEG 2000, 50 mM acetate buffer (pH 5.2) and 6.4% (w/v) PEG 200 or2.5% (w/v) PEG 1000, 50 mM acetate buffer (pH 4.8) and 25.7% (w/v) PEG 2000 MME Naegleria gruberi

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
1.2.1.12 additional information
-
additional information the enzyme presents allosteric positive cooperativity for substrates NAD+ and D-glyceraldehyde 3-phosphate, kinetic analysis, overview Naegleria gruberi

Natural Substrates/ Products (Substrates)

EC Number Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
1.2.1.12 D-glyceraldehyde 3-phosphate + phosphate + NAD+ Naegleria gruberi
-
3-phospho-D-glyceroyl phosphate + NADH + H+
-
?
1.2.1.107 D-glyceraldehyde 3-phosphate + arsenate + NAD+ Naegleria gruberi
-
1-arsono-3-phospho-D-glycerate + NADH + H+
-
ir

Organism

EC Number Organism UniProt Comment Textmining
1.2.1.12 Naegleria gruberi
-
nonpathogenic, free-living amoeba
-
1.2.1.107 Naegleria gruberi
-
-
-

Purification (Commentary)

EC Number Purification (Comment) Organism
1.2.1.12 recombinant His-tagged enzyme from Escherichia coli strain BL21(DE3) by nickell affinity chromatography, dialysis, and gel filtration Naegleria gruberi
1.2.1.107 nickel affinity column chromatography and Superdex 75 gel filtration Naegleria gruberi

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
1.2.1.12 D-glyceraldehyde 3-phosphate + phosphate + NAD+
-
Naegleria gruberi 3-phospho-D-glyceroyl phosphate + NADH + H+
-
?
1.2.1.107 D-glyceraldehyde 3-phosphate + arsenate + NAD+
-
Naegleria gruberi 1-arsono-3-phospho-D-glycerate + NADH + H+
-
ir

Synonyms

EC Number Synonyms Comment Organism
1.2.1.12 GAPDH
-
Naegleria gruberi
1.2.1.12 glyceraldehyde-3-phosphate dehydrogenase
-
Naegleria gruberi
1.2.1.107 GAPDH
-
Naegleria gruberi

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
1.2.1.12 25
-
-
Naegleria gruberi
1.2.1.107 25
-
-
Naegleria gruberi

Temperature Range [°C]

EC Number Temperature Minimum [°C] Temperature Maximum [°C] Comment Organism
1.2.1.12 10 35 activity range, profile overview Naegleria gruberi
1.2.1.107 20 30 high activity between 20 and 30°C Naegleria gruberi

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
1.2.1.12 8
-
-
Naegleria gruberi
1.2.1.107 8
-
-
Naegleria gruberi

pH Range

EC Number pH Minimum pH Maximum Comment Organism
1.2.1.12 6 10 activity range, profile overview Naegleria gruberi
1.2.1.107 7.5 8.5 high activity between pH 7.5 and 8.5 Naegleria gruberi

Cofactor

EC Number Cofactor Comment Organism Structure
1.2.1.12 NAD+
-
Naegleria gruberi
1.2.1.107 NAD+
-
Naegleria gruberi