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Literature summary extracted from

  • Moretti, R.; Thorson, J.S.
    Enhancing the latent nucleotide triphosphate flexibility of the glucose-1-phosphate thymidylyltransferase RmlA (2007), J. Biol. Chem., 282, 16942-16947.
    View publication on PubMed

Cloned(Commentary)

EC Number Cloned (Comment) Organism
2.7.7.24 expressed in Escherichia coli BL21 (DE3) cells Salmonella enterica typhimurium
2.7.7.24 expressed in Escherichia coli BL21(DE3) cells Salmonella enterica typhimurium

Protein Variants

EC Number Protein Variants Comment Organism
2.7.7.24 Q83A 0.2% relative activity compared to the wild type enzyme, using dTTP as a substrate Salmonella enterica typhimurium
2.7.7.24 Q83A reduced activity with dTTP and increased activity with dATP and ATP compared to the wild type enzyme Salmonella enterica typhimurium
2.7.7.24 Q83D 0.2% relative activity compared to the wild type enzyme, using dTTP as a substrate Salmonella enterica typhimurium
2.7.7.24 Q83D reduced activity with dTTP and increased activity with dGTP and GTP compared to the wild type enzyme Salmonella enterica typhimurium
2.7.7.24 Q83E 1.0% relative activity compared to the wild type enzyme, using dTTP as a substrate Salmonella enterica typhimurium
2.7.7.24 Q83E wild type activity with dTTP Salmonella enterica typhimurium
2.7.7.24 Q83N 0.3% relative activity compared to the wild type enzyme, using dTTP as a substrate Salmonella enterica typhimurium
2.7.7.24 Q83N reduced activity with dTTP and increased activity with dATP and ATP compared to the wild type enzyme Salmonella enterica typhimurium
2.7.7.24 Q83S 0.3% relative activity compared to the wild type enzyme, using dTTP as a substrate Salmonella enterica typhimurium
2.7.7.24 Q83S reduced activity with dTTP and increased activity with dATP and ATP compared to the wild type enzyme Salmonella enterica typhimurium

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
2.7.7.24 0.29
-
 dTTP wild type enzyme, apparent value, with 10 mM glucose-1-phosphate, 5 mM MgCl2, 10 units/ml inorganic pyrophosphatase, in 100 mM Tris-HCl, pH 8.0, at 37°C Salmonella enterica typhimurium
2.7.7.24 0.29
-
 dTTP apparent value, wild type enzyme, in 100 mM Tris-HCl, pH 8.0, at 37°C Salmonella enterica typhimurium
2.7.7.24 1.1
-
 dATP mutant enzyme Q83S, apparent value, with 10 mM glucose-1-phosphate, 5 mM MgCl2, 10 units/ml inorganic pyrophosphatase, in 100 mM Tris-HCl, pH 8.0, at 37°C Salmonella enterica typhimurium
2.7.7.24 1.1
-
 dATP apparent value, mutant enzyme Q83S, in 100 mM Tris-HCl, pH 8.0, at 37°C Salmonella enterica typhimurium
2.7.7.24 1.3
-
 GTP mutant enzyme Q83D, apparent value, with 10 mM glucose-1-phosphate, 5 mM MgCl2, 10 units/ml inorganic pyrophosphatase, in 100 mM Tris-HCl, pH 8.0, at 37°C Salmonella enterica typhimurium
2.7.7.24 1.3
-
 GTP apparent value, mutant enzyme Q83D, in 100 mM Tris-HCl, pH 8.0, at 37°C Salmonella enterica typhimurium
2.7.7.24 2.4
-
 dTTP mutant enzyme Q83S, apparent value, with 10 mM glucose-1-phosphate, 5 mM MgCl2, 10 units/ml inorganic pyrophosphatase, in 100 mM Tris-HCl, pH 8.0, at 37°C Salmonella enterica typhimurium
2.7.7.24 2.4
-
 dTTP apparent value, mutant enzyme Q83S, in 100 mM Tris-HCl, pH 8.0, at 37°C Salmonella enterica typhimurium
2.7.7.24 3.4
-
 ATP mutant enzyme Q83S, apparent value, with 10 mM glucose-1-phosphate, 5 mM MgCl2, 10 units/ml inorganic pyrophosphatase, in 100 mM Tris-HCl, pH 8.0, at 37°C Salmonella enterica typhimurium
2.7.7.24 3.4
-
 ATP apparent value, mutant enzyme Q83S, in 100 mM Tris-HCl, pH 8.0, at 37°C Salmonella enterica typhimurium
2.7.7.24 3.7
-
 dGTP wild type enzyme, apparent value, with 10 mM glucose-1-phosphate, 5 mM MgCl2, 10 units/ml inorganic pyrophosphatase, in 100 mM Tris-HCl, pH 8.0, at 37°C Salmonella enterica typhimurium
2.7.7.24 3.7
-
 dGTP apparent value, wild type enzyme, in 100 mM Tris-HCl, pH 8.0, at 37°C Salmonella enterica typhimurium
2.7.7.24 4.6
-
 UTP wild type enzyme, apparent value, with 10 mM glucose-1-phosphate, 5 mM MgCl2, 10 units/ml inorganic pyrophosphatase, in 100 mM Tris-HCl, pH 8.0, at 37°C Salmonella enterica typhimurium
2.7.7.24 4.6
-
 UTP apparent value, wild type enzyme, in 100 mM Tris-HCl, pH 8.0, at 37°C Salmonella enterica typhimurium
2.7.7.24 6.2
-
 dTTP mutant enzyme Q83D, apparent value, with 10 mM glucose-1-phosphate, 5 mM MgCl2, 10 units/ml inorganic pyrophosphatase, in 100 mM Tris-HCl, pH 8.0, at 37°C Salmonella enterica typhimurium
2.7.7.24 6.2
-
 dTTP apparent value, mutant enzyme Q83D, in 100 mM Tris-HCl, pH 8.0, at 37°C Salmonella enterica typhimurium
2.7.7.24 7.8
-
 dATP wild type enzyme, apparent value, with 10 mM glucose-1-phosphate, 5 mM MgCl2, 10 units/ml inorganic pyrophosphatase, in 100 mM Tris-HCl, pH 8.0, at 37°C Salmonella enterica typhimurium
2.7.7.24 7.8
-
 dATP apparent value, wild type enzyme, in 100 mM Tris-HCl, pH 8.0, at 37°C Salmonella enterica typhimurium
2.7.7.24 8.5
-
 GTP wild type enzyme, apparent value, with 10 mM glucose-1-phosphate, 5 mM MgCl2, 10 units/ml inorganic pyrophosphatase, in 100 mM Tris-HCl, pH 8.0, at 37°C Salmonella enterica typhimurium
2.7.7.24 8.5
-
 GTP apparent value, wild type enzyme, in 100 mM Tris-HCl, pH 8.0, at 37°C Salmonella enterica typhimurium
2.7.7.24 9
-
 ATP wild type enzyme, apparent value, with 10 mM glucose-1-phosphate, 5 mM MgCl2, 10 units/ml inorganic pyrophosphatase, in 100 mM Tris-HCl, pH 8.0, at 37°C Salmonella enterica typhimurium
2.7.7.24 9
-
 ATP apparent value, wild type enzyme, in 100 mM Tris-HCl, pH 8.0, at 37°C Salmonella enterica typhimurium
2.7.7.24 10.5
-
 dCTP wild type enzyme, apparent value, with 10 mM glucose-1-phosphate, 5 mM MgCl2, 10 units/ml inorganic pyrophosphatase, in 100 mM Tris-HCl, pH 8.0, at 37°C Salmonella enterica typhimurium
2.7.7.24 10.5
-
 dCTP apparent value, wild type enzyme, in 100 mM Tris-HCl, pH 8.0, at 37°C Salmonella enterica typhimurium
2.7.7.24 11.5
-
 dGTP mutant enzyme Q83D, apparent value, with 10 mM glucose-1-phosphate, 5 mM MgCl2, 10 units/ml inorganic pyrophosphatase, in 100 mM Tris-HCl, pH 8.0, at 37°C Salmonella enterica typhimurium
2.7.7.24 11.5
-
 dGTP apparent value, mutant enzyme Q83D, in 100 mM Tris-HCl, pH 8.0, at 37°C Salmonella enterica typhimurium
2.7.7.24 13.7
-
 CTP wild type enzyme, apparent value, with 10 mM glucose-1-phosphate, 5 mM MgCl2, 10 units/ml inorganic pyrophosphatase, in 100 mM Tris-HCl, pH 8.0, at 37°C Salmonella enterica typhimurium
2.7.7.24 13.7
-
 CTP apparent value, wild type enzyme, in 100 mM Tris-HCl, pH 8.0, at 37°C Salmonella enterica typhimurium

Organism

EC Number Organism UniProt Comment Textmining
2.7.7.24 Salmonella enterica typhimurium
-
-
-
2.7.7.24 Salmonella enterica typhimurium LT2
-
-
-

Purification (Commentary)

EC Number Purification (Comment) Organism
2.7.7.24 Ni-NTA agarose resin chromatography Salmonella enterica typhimurium
2.7.7.24 PD-10 gel filtration, nickel-nitrilotriacetic acid agarose resin or spin column chromatography Salmonella enterica typhimurium

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
2.7.7.24 ATP + alpha-D-glucose 1-phosphate
-
 Salmonella enterica typhimurium ADP-glucose + diphosphate
-
 ?
2.7.7.24 CTP + alpha-D-glucose 1-phosphate
-
 Salmonella enterica typhimurium CDP-glucose + diphosphate
-
 ?
2.7.7.24 dATP + alpha-D-glucose 1-phosphate
-
 Salmonella enterica typhimurium dADP-glucose + diphosphate
-
 ?
2.7.7.24 dATP + alpha-D-glucose 1-phosphate
-
 Salmonella enterica typhimurium LT2 dADP-glucose + diphosphate
-
 ?
2.7.7.24 dCTP + alpha-D-glucose 1-phosphate
-
 Salmonella enterica typhimurium dCDP-glucose + diphosphate
-
 ?
2.7.7.24 dCTP + alpha-D-glucose 1-phosphate
-
 Salmonella enterica typhimurium LT2 dCDP-glucose + diphosphate
-
 ?
2.7.7.24 dGTP + alpha-D-glucose 1-phosphate
-
 Salmonella enterica typhimurium dGDP-glucose + diphosphate
-
 ?
2.7.7.24 dGTP + alpha-D-glucose 1-phosphate
-
 Salmonella enterica typhimurium LT2 dGDP-glucose + diphosphate
-
 ?
2.7.7.24 dTTP + alpha-D-glucose 1-phosphate
-
 Salmonella enterica typhimurium dTDP-glucose + diphosphate
-
 ?
2.7.7.24 dTTP + alpha-D-glucose 1-phosphate
-
 Salmonella enterica typhimurium LT2 dTDP-glucose + diphosphate
-
 ?
2.7.7.24 GTP + alpha-D-glucose 1-phosphate
-
 Salmonella enterica typhimurium GDP-glucose + diphosphate
-
 ?
2.7.7.24 additional information utilizes all eight naturally occurring NTPs with varying levels of catalytic efficiency, even in the presence of nonnative sugar-1-phosphates Salmonella enterica typhimurium ?
-
 ?
2.7.7.24 additional information utilizes all eight naturally occurring NTPs with varying levels of catalytic efficiency, even in the presence of nonnative sugar-1-phosphates Salmonella enterica typhimurium LT2 ?
-
 ?
2.7.7.24 UTP + alpha-D-glucose 1-phosphate
-
 Salmonella enterica typhimurium UDP-glucose + diphosphate
-
 ?

Synonyms

EC Number Synonyms Comment Organism
2.7.7.24 RmlA
-
 Salmonella enterica typhimurium

Turnover Number [1/s]

EC Number Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
2.7.7.24 0.06
-
 ATP wild type enzyme, apparent value, with 10 mM glucose-1-phosphate, 5 mM MgCl2, 10 units/ml inorganic pyrophosphatase, in 100 mM Tris-HCl, pH 8.0, at 37°C Salmonella enterica typhimurium
2.7.7.24 0.06
-
 ATP apparent value, wild type enzyme, in 100 mM Tris-HCl, pH 8.0, at 37°C Salmonella enterica typhimurium
2.7.7.24 0.098
-
 CTP apparent value, wild type enzyme, in 100 mM Tris-HCl, pH 8.0, at 37°C Salmonella enterica typhimurium
2.7.7.24 0.1
-
 CTP wild type enzyme, apparent value, with 10 mM glucose-1-phosphate, 5 mM MgCl2, 10 units/ml inorganic pyrophosphatase, in 100 mM Tris-HCl, pH 8.0, at 37°C Salmonella enterica typhimurium
2.7.7.24 0.14
-
 GTP wild type enzyme, apparent value, with 10 mM glucose-1-phosphate, 5 mM MgCl2, 10 units/ml inorganic pyrophosphatase, in 100 mM Tris-HCl, pH 8.0, at 37°C Salmonella enterica typhimurium
2.7.7.24 0.14
-
 GTP apparent value, wild type enzyme, in 100 mM Tris-HCl, pH 8.0, at 37°C Salmonella enterica typhimurium
2.7.7.24 0.21
-
 GTP mutant enzyme Q83D, apparent value, with 10 mM glucose-1-phosphate, 5 mM MgCl2, 10 units/ml inorganic pyrophosphatase, in 100 mM Tris-HCl, pH 8.0, at 37°C Salmonella enterica typhimurium
2.7.7.24 0.21
-
 GTP apparent value, mutant enzyme Q83D, in 100 mM Tris-HCl, pH 8.0, at 37°C Salmonella enterica typhimurium
2.7.7.24 0.48
-
 ATP mutant enzyme Q83S, apparent value, with 10 mM glucose-1-phosphate, 5 mM MgCl2, 10 units/ml inorganic pyrophosphatase, in 100 mM Tris-HCl, pH 8.0, at 37°C Salmonella enterica typhimurium
2.7.7.24 0.48
-
 ATP apparent value, mutant enzyme Q83S, in 100 mM Tris-HCl, pH 8.0, at 37°C Salmonella enterica typhimurium
2.7.7.24 1.22
-
 dCTP wild type enzyme, apparent value, with 10 mM glucose-1-phosphate, 5 mM MgCl2, 10 units/ml inorganic pyrophosphatase, in 100 mM Tris-HCl, pH 8.0, at 37°C Salmonella enterica typhimurium
2.7.7.24 1.22
-
 dCTP apparent value, wild type enzyme, in 100 mM Tris-HCl, pH 8.0, at 37°C Salmonella enterica typhimurium
2.7.7.24 1.88
-
 dATP wild type enzyme, apparent value, with 10 mM glucose-1-phosphate, 5 mM MgCl2, 10 units/ml inorganic pyrophosphatase, in 100 mM Tris-HCl, pH 8.0, at 37°C Salmonella enterica typhimurium
2.7.7.24 1.88
-
 dATP apparent value, wild type enzyme, in 100 mM Tris-HCl, pH 8.0, at 37°C Salmonella enterica typhimurium
2.7.7.24 2.12
-
 dGTP wild type enzyme, apparent value, with 10 mM glucose-1-phosphate, 5 mM MgCl2, 10 units/ml inorganic pyrophosphatase, in 100 mM Tris-HCl, pH 8.0, at 37°C Salmonella enterica typhimurium
2.7.7.24 2.12
-
 dGTP apparent value, wild type enzyme, in 100 mM Tris-HCl, pH 8.0, at 37°C Salmonella enterica typhimurium
2.7.7.24 3.13
-
 dTTP mutant enzyme Q83D, apparent value, with 10 mM glucose-1-phosphate, 5 mM MgCl2, 10 units/ml inorganic pyrophosphatase, in 100 mM Tris-HCl, pH 8.0, at 37°C Salmonella enterica typhimurium
2.7.7.24 3.62
-
 dTTP apparent value, mutant enzyme Q83D, in 100 mM Tris-HCl, pH 8.0, at 37°C Salmonella enterica typhimurium
2.7.7.24 5
-
 dGTP mutant enzyme Q83D, apparent value, with 10 mM glucose-1-phosphate, 5 mM MgCl2, 10 units/ml inorganic pyrophosphatase, in 100 mM Tris-HCl, pH 8.0, at 37°C Salmonella enterica typhimurium
2.7.7.24 5
-
 dGTP apparent value, mutant enzyme Q83D, in 100 mM Tris-HCl, pH 8.0, at 37°C Salmonella enterica typhimurium
2.7.7.24 5.33
-
 dATP mutant enzyme Q83S, apparent value, with 10 mM glucose-1-phosphate, 5 mM MgCl2, 10 units/ml inorganic pyrophosphatase, in 100 mM Tris-HCl, pH 8.0, at 37°C Salmonella enterica typhimurium
2.7.7.24 5.33
-
 dATP apparent value, mutant enzyme Q83S, in 100 mM Tris-HCl, pH 8.0, at 37°C Salmonella enterica typhimurium
2.7.7.24 6.33
-
 dTTP mutant enzyme Q83S, apparent value, with 10 mM glucose-1-phosphate, 5 mM MgCl2, 10 units/ml inorganic pyrophosphatase, in 100 mM Tris-HCl, pH 8.0, at 37°C Salmonella enterica typhimurium
2.7.7.24 6.33
-
 dTTP apparent value, mutant enzyme Q83S, in 100 mM Tris-HCl, pH 8.0, at 37°C Salmonella enterica typhimurium
2.7.7.24 15
-
 dTTP wild type enzyme, apparent value, with 10 mM glucose-1-phosphate, 5 mM MgCl2, 10 units/ml inorganic pyrophosphatase, in 100 mM Tris-HCl, pH 8.0, at 37°C Salmonella enterica typhimurium
2.7.7.24 30.83
-
 dTTP apparent value, wild type enzyme, in 100 mM Tris-HCl, pH 8.0, at 37°C Salmonella enterica typhimurium
2.7.7.24 41
-
 UTP wild type enzyme, apparent value, with 10 mM glucose-1-phosphate, 5 mM MgCl2, 10 units/ml inorganic pyrophosphatase, in 100 mM Tris-HCl, pH 8.0, at 37°C Salmonella enterica typhimurium
2.7.7.24 41
-
 UTP apparent value, wild type enzyme, in 100 mM Tris-HCl, pH 8.0, at 37°C Salmonella enterica typhimurium