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Literature summary extracted from

  • Rapala-Kozik, M.; Olczak, M.; Ostrowska, K.; Starosta, A.; Kozik, A.
    Molecular characterization of thi3 gene involved in thiamine biosynthesis in Zea mays: cDNA sequence and enzymatic and structural properties of recombinant bifunctional protein with 4-amino-5-hydroxymethyl-2-methylpyrimidine (phosphate) kinase and thiamin (2007), Biochem. J., 408, 149-159.
    View publication on PubMedView publication on EuropePMC

Cloned(Commentary)

EC Number Cloned (Comment) Organism
2.5.1.3 expression in Escherichia coli Zea mays
2.7.1.49 expressed in Escherichia coli Zea mays

Protein Variants

EC Number Protein Variants Comment Organism
2.5.1.3 M134K phosphomethylpyrimidine kinase activity similar to wild-type, thiamin-phosphate diphosphorylase activity almost completely abolished Zea mays
2.5.1.3 M134K/T472D 83-97% residual phosphomethylpyrimidine kinase activity, 19-32% residual thiamin-phosphate diphosphorylase activity Zea mays
2.5.1.3 Q373L phosphomethylpyrimidine kinase activity similar to wild-type, 64-75% residual thiamin-phosphate diphosphorylase activity Zea mays
2.5.1.3 Q98L phosphomethylpyrimidine kinase activity similar to wild-type, thiamin-phosphate diphosphorylase activity almost completely abolished Zea mays
2.5.1.3 Q98L/Q373L phosphomethylpyrimidine kinase activity similar to wild-type, 7-13% residual thiamin-phosphate diphosphorylase activity Zea mays
2.5.1.3 S444A almost complete loss of phosphomethylpyrimidine kinase activity, 80-91% residual thiamin-phosphate diphosphorylase activity Zea mays
2.5.1.3 T472D phosphomethylpyrimidine kinase activity similar to wild-type, 46-54% residual thiamin-phosphate diphosphorylase activity Zea mays
2.7.1.49 M134K detrimental to HMP kinase activity Zea mays
2.7.1.49 M134K/T472D double mutant with 25% activity compared to the wild type enzyme Zea mays
2.7.1.49 Q373L 30% reduced HMP kinase activity Zea mays
2.7.1.49 Q98L detrimental to HMP kinase activity Zea mays
2.7.1.49 Q98L/Q373L double mutant with 10% activity compared to the wild type enzyme Zea mays
2.7.1.49 S444A without any effect on activity Zea mays
2.7.1.49 T472D 50% reduced HMP kinase activity Zea mays

Inhibitors

EC Number Inhibitors Comment Organism Structure
2.5.1.3 4-amino-5-hydroxymethyl-2-methylpyrimidine diphosphate uncompetitive Zea mays
2.5.1.3 ATP uncompetitive Zea mays

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
2.5.1.3 0.0085
-
 4-methyl-5-(2-hydroxyethyl)thiazole phosphate
-
 Zea mays
2.5.1.3 0.0128
-
 4-amino-5-hydroxymethyl-2-methylpyrimidine diphosphate 37°C, pH 8.0 Zea mays

Metals/Ions

EC Number Metals/Ions Comment Organism Structure
2.5.1.3 Mg2+ required Zea mays

Molecular Weight [Da]

EC Number Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
2.5.1.3 55000
-
 2 * 57800, calculated, 2 * 55000, SDS-PAGE Zea mays
2.5.1.3 57800
-
 2 * 57800, calculated, 2 * 55000, SDS-PAGE Zea mays
2.5.1.3 95000
-
 gel filtration Zea mays
2.7.1.49 47500
-
 2 * 47500, gel filtration Zea mays
2.7.1.49 57800
-
 calculated from amino acid sequence Zea mays
2.7.1.49 95000
-
 gel filtration Zea mays

Organism

EC Number Organism UniProt Comment Textmining
2.5.1.3 Zea mays Q2UVH9 isoform Thi3, bifunctional phosphomethylpyrimidine kinase/thiamin-phosphate diphosphorylase
-
2.7.1.49 Zea mays
-
-
-

Purification (Commentary)

EC Number Purification (Comment) Organism
2.7.1.49 Superdex 200 HR 10/50 gel filtration Zea mays

Source Tissue

EC Number Source Tissue Comment Organism Textmining
2.7.1.49 leaf
-
 Zea mays
-

Specific Activity [micromol/min/mg]

EC Number Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
2.5.1.3 0.05
-
 37°C, pH 8.0, presence of ATP Zea mays
2.5.1.3 0.52
-
 37°C, pH 8.0 Zea mays
2.5.1.3 2.46
-
 37°C, pH 8.0, presence of Mg2+ Zea mays

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
2.5.1.3 4-amino-5-hydroxymethyl-2-methylpyrimidine diphosphate + 4-methyl-5-(2-hydroxyethyl)thiazole phosphate
-
 Zea mays thiamine monophosphate + diphosphate + phosphate
-
 ?
2.5.1.3 additional information substrate 4-amino-5-hydroxymethyl-2-methylpyrimidine diphosphate cannot be replaced by 4-amino-5-hydroxymethyl-2-methylpyrimidine phosphate or 4-amino-5-hydroxymethyl-2-methylpyrimidine, nor substrate 4-methyl-5-(2-hydroxyethyl)thiazole phosphate by 4-methyl-5-(2-hydroxyethyl)thiazole Zea mays ?
-
 ?
2.7.1.49 ATP + 2-methyl-4-amino-5-hydroxymethylpyrimidine
-
 Zea mays ADP + 2-methyl-4-amino-5-phosphomethylpyrimidine
-
 ?

Subunits

EC Number Subunits Comment Organism
2.5.1.3 dimer 2 * 57800, calculated, 2 * 55000, SDS-PAGE Zea mays
2.5.1.3 More the activities of bifunctional phosphomethylpyrimidine kinase/thiamin-phosphate diphosphorylase are located in two distinct, N-terminal kinase and C-terminal synthase, domains Zea mays
2.7.1.49 dimer 2 * 47500, gel filtration Zea mays

Synonyms

EC Number Synonyms Comment Organism
2.7.1.49 4-amino-5-hydroxymethyl-2-methylpyrimidine (phosphate) kinase
-
 Zea mays
2.7.1.49 HMP kinase
-
 Zea mays
2.7.1.49 THI3 the enzyme shows two essentially different enzymatic activities of 4-amino-5-hydroxymethyl-2-methylpyrimidne (phosphate) kinase and thiamine monophosphate synthase Zea mays

Cofactor

EC Number Cofactor Comment Organism Structure
2.7.1.49 ATP
-
 Zea mays

Ki Value [mM]

EC Number Ki Value [mM] Ki Value maximum [mM] Inhibitor Comment Organism Structure
2.5.1.3 0.0034
-
 ATP
-
 Zea mays
2.5.1.3 0.0148
-
 4-amino-5-hydroxymethyl-2-methylpyrimidine diphosphate 37°C, pH 8.0 Zea mays

pI Value

EC Number Organism Comment pI Value Maximum pI Value
2.5.1.3 Zea mays calculated
-
 8.5
2.7.1.49 Zea mays calculated from amino acid sequence
-
 8.51