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Literature summary for 2.5.1.3 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
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Cloned(Commentary)

Cloned (Comment) Organism
expression in Escherichia coli Zea mays

Protein Variants

Protein Variants Comment Organism
M134K phosphomethylpyrimidine kinase activity similar to wild-type, thiamin-phosphate diphosphorylase activity almost completely abolished Zea mays
M134K/T472D 83-97% residual phosphomethylpyrimidine kinase activity, 19-32% residual thiamin-phosphate diphosphorylase activity Zea mays
Q373L phosphomethylpyrimidine kinase activity similar to wild-type, 64-75% residual thiamin-phosphate diphosphorylase activity Zea mays
Q98L phosphomethylpyrimidine kinase activity similar to wild-type, thiamin-phosphate diphosphorylase activity almost completely abolished Zea mays
Q98L/Q373L phosphomethylpyrimidine kinase activity similar to wild-type, 7-13% residual thiamin-phosphate diphosphorylase activity Zea mays
S444A almost complete loss of phosphomethylpyrimidine kinase activity, 80-91% residual thiamin-phosphate diphosphorylase activity Zea mays
T472D phosphomethylpyrimidine kinase activity similar to wild-type, 46-54% residual thiamin-phosphate diphosphorylase activity Zea mays

Inhibitors

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

KM Value [mM]

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

Metals/Ions

Metals/Ions Comment Organism Structure
Mg2+ required Zea mays

Molecular Weight [Da]

Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
55000
-
 2 * 57800, calculated, 2 * 55000, SDS-PAGE Zea mays
57800
-
 2 * 57800, calculated, 2 * 55000, SDS-PAGE Zea mays
95000
-
 gel filtration Zea mays

Organism

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

Specific Activity [micromol/min/mg]

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

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
4-amino-5-hydroxymethyl-2-methylpyrimidine diphosphate + 4-methyl-5-(2-hydroxyethyl)thiazole phosphate
-
 Zea mays thiamine monophosphate + diphosphate + phosphate
-
 ?
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 ?
-
 ?

Subunits

Subunits Comment Organism
dimer 2 * 57800, calculated, 2 * 55000, SDS-PAGE Zea mays
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

Ki Value [mM]

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

pI Value

Organism Comment pI Value Maximum pI Value
Zea mays calculated
-
 8.5