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Literature summary extracted from

  • Reddick, J.J.; Kinsland, C.; Nicewonger, R.; Christian, T.; Downs, D.M.; Winkler, M.E.; Begley, T.P.
    Overexpression, purification and characterization of two pyrimidine kinases involved in the biosynthesis of thiamin: 4-amino-5-hydroxymethyl-2-methylpyrimidine kinase and 4-amino-5-hydroxymethyl-2-methylpyrimidine phosphate kinase (1998), Tetrahedron, 54, 15983-15991.
No PubMed abstract available

Application

EC Number Application Comment Organism
2.7.1.49 synthesis useful reagent for the preparation of intermediates on the thiamin biosynthetic pathway Escherichia coli

Cloned(Commentary)

EC Number Cloned (Comment) Organism
2.7.1.49 pET overexpression system Escherichia coli
2.7.4.7 overexpression of ThiD in Escherichia coli Escherichia coli

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
2.7.1.49 0.0285
-
 pyridoxine pH 8.0, 25°C Escherichia coli
2.7.1.49 0.0335
-
 2-methyl-4-amino-5-hydroxymethylpyrimidine pH 8.0, 25°C Escherichia coli
2.7.1.49 0.1431
-
 ATP pH 8.0, 25°C Escherichia coli
2.7.4.7 0.023
-
 2-methyl-4-amino-5-hydroxymethylpyrimidine phosphate pH 8.0, 25°C, recombinant ThiD Escherichia coli
2.7.4.7 0.15
-
 2-methyl-4-amino-5-hydroxymethylpyrimidine pH 8.0, 25°C, recombinant ThiD Escherichia coli
2.7.4.7 0.193
-
 ATP pH 8.0, 25°C, recombinant ThiD Escherichia coli

Molecular Weight [Da]

EC Number Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
2.7.1.49 additional information
-
 gel filtration indicates the enzyme is tetrameric in the native state Escherichia coli
2.7.1.49 33000
-
 4 * 33000, SDS-PAGE Escherichia coli
2.7.1.49 33534
-
 4 * 33534, predicted from amino acid sequence Escherichia coli
2.7.4.7 31232
-
 4 * 31232, deduced from nucleotide sequence Escherichia coli
2.7.4.7 33000
-
 4 * 33000, SDS-PAGE Escherichia coli

Natural Substrates/ Products (Substrates)

EC Number Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
2.7.1.49 ATP + 2-methyl-4-amino-5-hydroxymethylpyrimidine Escherichia coli involved in thiamine biosynthetic pathway, first steps ADP + 2-methyl-4-amino-5-phosphomethylpyrimidine
-
 ?
2.7.4.7 ATP + 2-methyl-4-amino-5-hydroxymethylpyrimidine Escherichia coli involved in thiamine biosynthesis ADP + 2-methyl-4-amino-5-phosphomethylpyrimidine
-
 ?
2.7.4.7 ATP + 2-methyl-4-amino-5-phosphomethylpyrimidine Escherichia coli
-
 ADP + 2-methyl-4-amino-5-diphosphomethylpyrimidine
-
 ?

Organism

EC Number Organism UniProt Comment Textmining
2.7.1.49 Escherichia coli
-
 BL21(DE3)
-
2.7.4.7 Escherichia coli
-
-
-

Purification (Commentary)

EC Number Purification (Comment) Organism
2.7.1.49
-
 Escherichia coli
2.7.4.7 his-tagged recombinant ThiD Escherichia coli

Storage Stability

EC Number Storage Stability Organism
2.7.1.49 -70°C, stored in 10% glycerol, loses half of its activity after 1 month Escherichia coli

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
2.7.1.49 ATP + 2-methyl-4-amino-5-hydroxymethylpyrimidine
-
 Escherichia coli ADP + 2-methyl-4-amino-5-phosphomethylpyrimidine
-
 ?
2.7.1.49 ATP + 2-methyl-4-amino-5-hydroxymethylpyrimidine involved in thiamine biosynthetic pathway, first steps Escherichia coli ADP + 2-methyl-4-amino-5-phosphomethylpyrimidine
-
 ?
2.7.1.49 ATP + pyridoxine
-
 Escherichia coli ADP + pyridoxine 5'-phosphate
-
 ?
2.7.1.49 additional information enzyme is probably different from previously isolated HMP kinase, because the subunit molecular masses are significantly different Escherichia coli ?
-
 ?
2.7.4.7 ATP + 2-methyl-4-amino-5-hydroxymethylpyrimidine
-
 Escherichia coli ADP + 2-methyl-4-amino-5-phosphomethylpyrimidine
-
 ?
2.7.4.7 ATP + 2-methyl-4-amino-5-hydroxymethylpyrimidine involved in thiamine biosynthesis Escherichia coli ADP + 2-methyl-4-amino-5-phosphomethylpyrimidine
-
 ?
2.7.4.7 ATP + 2-methyl-4-amino-5-phosphomethylpyrimidine
-
 Escherichia coli ADP + 2-methyl-4-amino-5-diphosphomethylpyrimidine
-
 ?

Subunits

EC Number Subunits Comment Organism
2.7.1.49 tetramer 4 * 33000, SDS-PAGE Escherichia coli
2.7.1.49 tetramer 4 * 33534, predicted from amino acid sequence Escherichia coli
2.7.4.7 tetramer 4 * 33000, SDS-PAGE Escherichia coli
2.7.4.7 tetramer 4 * 31232, deduced from nucleotide sequence Escherichia coli

Synonyms

EC Number Synonyms Comment Organism
2.7.4.7 HMP-P kinase
-
 Escherichia coli
2.7.4.7 ThiD
-
 Escherichia coli

Turnover Number [1/s]

EC Number Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
2.7.1.49 0.36
-
 pyridoxine pH 8.0, 25°C Escherichia coli
2.7.1.49 0.43
-
 ATP pH 8.0, 25°C Escherichia coli
2.7.1.49 0.43
-
 2-methyl-4-amino-5-hydroxymethylpyrimidine pH 8.0, 25°C Escherichia coli
2.7.4.7 0.07
-
 ATP pH 8.0, 25°C, recombinant ThiD Escherichia coli
2.7.4.7 0.07
-
 2-methyl-4-amino-5-hydroxymethylpyrimidine phosphate pH 8.0, 25°C, recombinant ThiD Escherichia coli
2.7.4.7 0.0867
-
 2-methyl-4-amino-5-hydroxymethylpyrimidine pH 8.0, 25°C, recombinant ThiD Escherichia coli