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Literature summary extracted from

  • Kolappan, S.; Shen, D.L.; Mosi, R.; Sun, J.; McEachern, E.J.; Vocadlo, D.J.; Craig, L.
    Structures of lactate dehydrogenase A (LDHA) in apo, ternary and inhibitor-bound forms (2015), Acta Crystallogr. Sect. D, 71, 185-195.
    View publication on PubMed

Cloned(Commentary)

EC Number Cloned (Comment) Organism
1.1.1.27 recombinant expression of C-terminally His6-tagged enzyme from pET-29b-LDHA vector in Escherichia coli strain BL21(lambdaDE3) Homo sapiens

Crystallization (Commentary)

EC Number Crystallization (Comment) Organism
1.1.1.27 lactate dehydrogenase A in apo form, in ternary complex with oxalate and kanamycin, and in inhibitor-bound form, hanging drop vapour diffusion method, apo-LDHA crystals are grown by mixing of 0.002 ml of 25 mg/ml enzyme protein with 0.002 ml reservoir solution consisting of 100 mM Bis-Tris propane, pH 7.0, 20% v/v PEG 400, and 100 mM LiCl. LDHA-NADH crystals are grown by first incubating 25 mg/ml enzyme protein with 3 mM NADH for 2 h at 4°C and then setting up 0.004 ml drops with a 2:1:1 volume ratio of LDHA-NADH, reservoir solution, containing 18% w/v PEG 3350, and 50 mM HEPES, pH 6.8, and another reagent containing 0.16% w/v L-citrulline, 0.16% w/v L-ornithine hydrochloride, 0.16% w/v urea, 0.16% w/v oxalic acid, 0.16% w/v kanamycin monosulfate, and 0.16% w/v L-arginine in 0.02 M HEPES sodium, pH 6.8, 20°C. Apo and NADH-bound LDHA crystals appear after three weeks. LDHA-inhibitor complex crystals are obtained by adding 0.010 ml soaking solution containing 20 mM inhibitor, 100 mM HEPES, pH 7.5, 50 mM LiCl, 100 mM bis-tris propane pH 7.0, 20% v/v DMSO, and 25% PEG 8000, to 0.002 ml hanging drops containing apo LDHA crystals and allowing the crystals to sit at room temperature for 24 h, X-ray diffraction structure determination and analysis at 2.1-3.2 A resolution, molecular replacement method using model structure PDB ID1i10 Homo sapiens

Inhibitors

EC Number Inhibitors Comment Organism Structure
1.1.1.27 3-([3-carbamoyldimethoxypyrimidin-7-(2,4-dimethoxypyrimidin-5-yl)quinolin-4-yl]amino)benzoic acid enzyme binding structure, overview Homo sapiens
1.1.1.27 3-[7-(2,4-dimethoxypyrimidin-5-yl)-3-sulfamoylquinolin-4-yl]aminobenzoic acid enzyme binding structure, overview Homo sapiens

Natural Substrates/ Products (Substrates)

EC Number Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
1.1.1.27 (S)-lactate + NAD+ Homo sapiens
-
 pyruvate + NADH + H+
-
 r

Organism

EC Number Organism UniProt Comment Textmining
1.1.1.27 Homo sapiens P00338
-
-

Purification (Commentary)

EC Number Purification (Comment) Organism
1.1.1.27 recombinant C-terminally His6-tagged enzyme from Escherichia coli strain BL21(lambdaDE3) by centrfugation at 40000 g, nickel affinity chromatography, dialysis and ultrafiltration, followed by gel filtration, to over 95% purity Homo sapiens

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
1.1.1.27 (S)-lactate + NAD+
-
 Homo sapiens pyruvate + NADH + H+
-
 r

Subunits

EC Number Subunits Comment Organism
1.1.1.27 More LDHA ternary structures, overview Homo sapiens
1.1.1.27 tetramer crystal structure analysis Homo sapiens

Synonyms

EC Number Synonyms Comment Organism
1.1.1.27 lactate dehydrogenase A
-
 Homo sapiens
1.1.1.27 LdhA
-
 Homo sapiens

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
1.1.1.27 37
-
 assay at, pyruvate reduction Homo sapiens

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
1.1.1.27 7.5
-
 assay at, pyruvate reduction Homo sapiens

Cofactor

EC Number Cofactor Comment Organism Structure
1.1.1.27 NAD+
-
 Homo sapiens
1.1.1.27 NADH
-
 Homo sapiens

Ki Value [mM]

EC Number Ki Value [mM] Ki Value maximum [mM] Inhibitor Comment Organism Structure
1.1.1.27 0.00059
-
 3-[7-(2,4-dimethoxypyrimidin-5-yl)-3-sulfamoylquinolin-4-yl]aminobenzoic acid pH 7.5, 37°C, recombinant His-tagged enzyme, pyruvate reduction, competitive versus NADH Homo sapiens
1.1.1.27 0.00252
-
 3-[7-(2,4-dimethoxypyrimidin-5-yl)-3-sulfamoylquinolin-4-yl]aminobenzoic acid pH 7.5, 37°C, recombinant His-tagged enzyme, pyruvate reduction, noncompetitive versus pyruvate Homo sapiens

IC50 Value

EC Number IC50 Value IC50 Value Maximum Comment Organism Inhibitor Structure
1.1.1.27 0.000039
-
 pH 7.5, 37°C, recombinant His-tagged enzyme, L-lactate oxidation Homo sapiens 3-[7-(2,4-dimethoxypyrimidin-5-yl)-3-sulfamoylquinolin-4-yl]aminobenzoic acid
1.1.1.27 0.00051
-
 pH 7.5, 37°C, recombinant His-tagged enzyme, L-lactate oxidation Homo sapiens 3-([3-carbamoyldimethoxypyrimidin-7-(2,4-dimethoxypyrimidin-5-yl)quinolin-4-yl]amino)benzoic acid
1.1.1.27 0.0022
-
 pH 7.5, 37°C, recombinant His-tagged enzyme, pyruvate reduction Homo sapiens 3-[7-(2,4-dimethoxypyrimidin-5-yl)-3-sulfamoylquinolin-4-yl]aminobenzoic acid
1.1.1.27 0.0144
-
 pH 7.5, 37°C, recombinant His-tagged enzyme, pyruvate reduction Homo sapiens 3-([3-carbamoyldimethoxypyrimidin-7-(2,4-dimethoxypyrimidin-5-yl)quinolin-4-yl]amino)benzoic acid