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Literature summary for 2.5.1.112 extracted from

  • Abe, I.; Tanaka, H.; Abe, T.; Noguchi, H.
    Enzymatic formation of unnatural cytokinin analogs by adenylate isopentenyltransferase from mulberry (2007), Biochem. Biophys. Res. Commun., 355, 795-800.
    View publication on PubMed
Search for more literature for 2.5.1.112

Cloned(Commentary)

Cloned (Comment) Organism
expressed in Escherichia coli BL21(DE3)pLysS cells Morus alba

Protein Variants

Protein Variants Comment Organism
D49A the mutant almost completely loses the enzyme activity for the prenyl transfer from dimethylallyl diphosphate to ADP Morus alba
F120A the mutant almost completely loses the enzyme activity for the prenyl transfer from dimethylallyl diphosphate to ADP Morus alba
F157A the mutant almost completely loses the enzyme activity for the prenyl transfer from dimethylallyl diphosphate to ADP Morus alba
F93A the mutant almost completely loses the enzyme activity for the prenyl transfer from dimethylallyl diphosphate to ADP Morus alba
Q255A the mutant almost completely loses the enzyme activity for the prenyl transfer from dimethylallyl diphosphate to ADP Morus alba
W159A the mutant almost completely loses the enzyme activity for the prenyl transfer from dimethylallyl diphosphate to ADP Morus alba
Y153A the mutant almost completely loses the enzyme activity for the prenyl transfer from dimethylallyl diphosphate to ADP Morus alba
Y170A the mutant almost completely loses the enzyme activity for the prenyl transfer from dimethylallyl diphosphate to ADP Morus alba
Y170A the mutant retains 84% of wild type activity Morus alba
Y217A the mutant almost completely loses the enzyme activity for the prenyl transfer from dimethylallyl diphosphate to ADP Morus alba
Y54A the mutant almost completely loses the enzyme activity for the prenyl transfer from dimethylallyl diphosphate to ADP Morus alba

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.0374
-
 dimethylallyl diphosphate in 100 mM Tris-HCl buffer, pH 7.0, containing 50 mM KCl, 2 mM MnCl2, 1 mg/ml bovine serum albumin, and 1 mM dithiohtreitol, at 25°C Morus alba
0.0682
-
 ADP in 100 mM Tris-HCl buffer, pH 7.0, containing 50 mM KCl, 2 mM MnCl2, 1 mg/ml bovine serum albumin, and 1 mM dithiohtreitol, at 25°C Morus alba

Metals/Ions

Metals/Ions Comment Organism Structure
Mg2+ the enzyme is dependent on presence of divalent cation, and addition of 2 mM MnCl2 is 1.3times more effective than that of 10 mM MgCl2 Morus alba
Mn2+ the enzyme is dependent on presence of divalent cation, and addition of 2 mM MnCl2 is 1.3times more effective than that of 10 mM MgCl2 Morus alba

Molecular Weight [Da]

Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
35810
-
 x * 35810, calculated from amino acid sequence Morus alba

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
dimethylallyl diphosphate + ADP Morus alba 100% activity diphosphate + N6-(dimethylallyl)adenosine 5'-diphosphate
-
 ?
dimethylallyl diphosphate + AMP Morus alba 6.6% activity compared to ADP diphosphate + N6-(dimethylallyl)adenosine phosphate
-
 ?
dimethylallyl diphosphate + ATP Morus alba 82% activity compared to ADP diphosphate + N6-(dimethylallyl)adenosine 5'-triphosphate
-
 ?

Organism

Organism UniProt Comment Textmining
Morus alba Q08ET4
-
-

Purification (Commentary)

Purification (Comment) Organism
Ni-chelate affinity column chromatography Morus alba

Source Tissue

Source Tissue Comment Organism Textmining
leaf
-
 Morus alba
-

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
4-hydroxy-3-methyl-2-(E)-butenyl diphosphate + ADP 2.2% activity compared to dimethylallyl diphosphate and ADP Morus alba trans-zeatin riboside 5'-diphosphate + diphosphate
-
 ?
4-hydroxy-3-methyl-2-(E)-butenyl diphosphate + ATP 1.4% activity compared to dimethylallyl diphosphate and ADP Morus alba trans-zeatin riboside 5'-triphosphate + diphosphate
-
 ?
dimethylallyl diphosphate + ADP 100% activity Morus alba diphosphate + N6-(dimethylallyl)adenosine 5'-diphosphate
-
 ?
dimethylallyl diphosphate + AMP 6.6% activity compared to ADP Morus alba diphosphate + N6-(dimethylallyl)adenosine phosphate
-
 ?
dimethylallyl diphosphate + ATP 82% activity compared to ADP Morus alba diphosphate + N6-(dimethylallyl)adenosine 5'-triphosphate
-
 ?
dimethylallyl diphosphate + CDP 30% activity compared to ADP Morus alba ?
-
 ?
dimethylallyl diphosphate + dADP 50% activity compared to ADP Morus alba ?
-
 ?
dimethylallyl diphosphate + dATP 36% activity compared to ADP Morus alba ?
-
 ?
dimethylallyl diphosphate + GDP 6.4% activity compared to ADP Morus alba ?
-
 ?
geranyl diphosphate + ADP 1.2% activity compared to dimethylallyl diphosphate and ADP Morus alba ?
-
 ?
isopentenyl diphosphate + ADP 0.3% activity compared to dimethylallyl diphosphate and ADP Morus alba ?
-
 ?
additional information no activity with farnesyl diphosphate, dAMP, CMP, GMP, IMP, IDP, UMP, UDP, FAD and NADH Morus alba ?
-
 ?

Subunits

Subunits Comment Organism
? x * 35810, calculated from amino acid sequence Morus alba

Synonyms

Synonyms Comment Organism
adenylate isopentenyltransferase
-
 Morus alba
AIPT
-
 Morus alba

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
0.024
-
 ADP in 100 mM Tris-HCl buffer, pH 7.0, containing 50 mM KCl, 2 mM MnCl2, 1 mg/ml bovine serum albumin, and 1 mM dithiohtreitol, at 25°C Morus alba
0.035
-
 dimethylallyl diphosphate in 100 mM Tris-HCl buffer, pH 7.0, containing 50 mM KCl, 2 mM MnCl2, 1 mg/ml bovine serum albumin, and 1 mM dithiohtreitol, at 25°C Morus alba

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
6 7
-
 Morus alba

Cofactor

Cofactor Comment Organism Structure
ADP preferred cofactor Morus alba
ATP
-
 Morus alba

kcat/KM [mM/s]

kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
0.344
-
 ADP in 100 mM Tris-HCl buffer, pH 7.0, containing 50 mM KCl, 2 mM MnCl2, 1 mg/ml bovine serum albumin, and 1 mM dithiohtreitol, at 25°C Morus alba
0.921
-
 dimethylallyl diphosphate in 100 mM Tris-HCl buffer, pH 7.0, containing 50 mM KCl, 2 mM MnCl2, 1 mg/ml bovine serum albumin, and 1 mM dithiohtreitol, at 25°C Morus alba