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Literature summary extracted from

  • Frebortova, J.; Greplova, M.; Seidl, M.F.; Heyl, A.; Frebort, I.
    Biochemical characterization of putative adenylate dimethylallyltransferase and cytokinin dehydrogenase from Nostoc sp. PCC 7120 (2015), PLoS ONE, 10, e0138468.
    View publication on PubMedView publication on EuropePMC

Cloned(Commentary)

EC Number Cloned (Comment) Organism
2.5.1.27 expressed in Escherichia coli Top10 cells Nostoc sp.
2.5.1.27 gene NoIPT1, DNA and amino acid sequence determination and analysis, phylogenetic analysis, functional recombinant expression of His-tagged enzyme in Escherichia coli strain BL21 (DE3) STAR Nostoc sp.

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
2.5.1.27 additional information
-
 additional information Michaelis-Menten kinetics Nostoc sp.
2.5.1.27 0.0063
-
 AMP at pH 7.5 and 25°C Nostoc sp.
2.5.1.27 0.02
-
 dimethylallyl diphosphate pH 7.5, 25°C, recombinant enzyme Nostoc sp.
2.5.1.27 0.02705
-
 dimethylallyl diphosphate at pH 7.5 and 25°C Nostoc sp.
2.5.1.27 0.15
-
 AMP pH 7.5, 25°C, recombinant enzyme Nostoc sp.

Metals/Ions

EC Number Metals/Ions Comment Organism Structure
2.5.1.27 Mg2+ required Nostoc sp.

Molecular Weight [Da]

EC Number Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
2.5.1.27 27900
-
-
 Nostoc sp.
2.5.1.27 28300
-
-
 Nostoc sp.

Natural Substrates/ Products (Substrates)

EC Number Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
2.5.1.27 dimethylallyl diphosphate + AMP Nostoc sp.
-
 diphosphate + N6-(dimethylallyl)adenosine 5'-phosphate
-
 ?
2.5.1.27 dimethylallyl diphosphate + AMP Nostoc sp. PCC 7120
-
 diphosphate + N6-(dimethylallyl)adenosine 5'-phosphate
-
 ?

Organism

EC Number Organism UniProt Comment Textmining
2.5.1.27 Nostoc sp.
-
-
-
2.5.1.27 Nostoc sp. Q8Z078 gene NoIPT1
-
2.5.1.27 Nostoc sp. PCC 7120
-
-
-
2.5.1.27 Nostoc sp. PCC 7120 Q8Z078 gene NoIPT1
-

Purification (Commentary)

EC Number Purification (Comment) Organism
2.5.1.27 Ni-NTA column chromatography Nostoc sp.
2.5.1.27 recombinant His-tagged enzyme from Escherichia coli strain BL21 (DE3) STAR by anion exchange and nickel affinity chromatography to apparent homogeneity Nostoc sp.

Specific Activity [micromol/min/mg]

EC Number Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
2.5.1.27 0.01122
-
 purified recombinant enzyme, pH 7.5, 25°C Nostoc sp.

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
2.5.1.27 (E)-4-hydroxy-3-methyl-but-2-enyl diphosphate + AMP HMBPP, the enzyme uses the hydroxylated side chain precursor to form trans-zeatin-5'-monophosphate in the reaction with AMP, reaction rate is 0.5% of the rate with dimethylallyl díphosphate Nostoc sp. diphosphate + trans-zeatin riboside 5'phosphate
-
 ?
2.5.1.27 (E)-4-hydroxy-3-methyl-but-2-enyl diphosphate + AMP HMBPP, the enzyme uses the hydroxylated side chain precursor to form trans-zeatin-5'-monophosphate in the reaction with AMP, reaction rate is 0.5% of the rate with dimethylallyl díphosphate Nostoc sp. PCC 7120 diphosphate + trans-zeatin riboside 5'phosphate
-
 ?
2.5.1.27 dimethylallyl diphosphate + AMP
-
 Nostoc sp. diphosphate + N6-(dimethylallyl)adenosine 5'-phosphate
-
 ?
2.5.1.27 dimethylallyl diphosphate + AMP
-
 Nostoc sp. PCC 7120 diphosphate + N6-(dimethylallyl)adenosine 5'-phosphate
-
 ?
2.5.1.27 additional information less than 0.5% activity with (E)-4-hydroxy-3-methyl-but-2-enyl diphosphate plus AMP Nostoc sp. ?
-
 ?
2.5.1.27 additional information no activity with ADP and ATP. Isopentenyl diphosphate, an isomer of DMAPP, is no substrate with either of the side chain acceptor substrates. The enzyme does not show tRNA IPT activity, as it does not prenylate synthetic tRNA-stem-loop minihelix substrate Nostoc sp. ?
-
 ?
2.5.1.27 additional information the enzyme shows no activity with ATP and ADP and no tRNA isopentenyl transferase activity Nostoc sp. ?
-
 ?
2.5.1.27 additional information less than 0.5% activity with (E)-4-hydroxy-3-methyl-but-2-enyl diphosphate plus AMP Nostoc sp. PCC 7120 ?
-
 ?
2.5.1.27 additional information the enzyme shows no activity with ATP and ADP and no tRNA isopentenyl transferase activity Nostoc sp. PCC 7120 ?
-
 ?
2.5.1.27 additional information no activity with ADP and ATP. Isopentenyl diphosphate, an isomer of DMAPP, is no substrate with either of the side chain acceptor substrates. The enzyme does not show tRNA IPT activity, as it does not prenylate synthetic tRNA-stem-loop minihelix substrate Nostoc sp. PCC 7120 ?
-
 ?

Subunits

EC Number Subunits Comment Organism
2.5.1.27 ? x * 27900, recombinant enzyme, SDS-PAGE, x * 28300, about, sequence calculation Nostoc sp.
2.5.1.27 ? x * 27900, SDS-PAGE Nostoc sp.
2.5.1.27 ? x * 28300, calculated from amino acid sequence Nostoc sp.

Synonyms

EC Number Synonyms Comment Organism
2.5.1.27 adenylate dimethylallyltransferase
-
 Nostoc sp.
2.5.1.27 adenylate IPT
-
 Nostoc sp.
2.5.1.27 adenylate isopentenyl transferase
-
 Nostoc sp.
2.5.1.27 adenylate-type isopentenyl transferase
-
 Nostoc sp.
2.5.1.27 IPT1
-
 Nostoc sp.
2.5.1.27 isopentenyl transferase
-
 Nostoc sp.
2.5.1.27 NoIPT1
-
 Nostoc sp.

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
2.5.1.27 25
-
 assay at Nostoc sp.

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
2.5.1.27 7.5
-
-
 Nostoc sp.

General Information

EC Number General Information Comment Organism
2.5.1.27 evolution adenylate-type isopentenyl transferases might have evolved several times independently, phylogenetic analysis. Key enzymes for cytokinin synthesis and degradation (adenylate IPT and CKX) are found only in few species of cyanobacteria Nostoc sp.
2.5.1.27 metabolism the biosynthesis of cytokinins starts with a transfer of dimethylallyl chain from dimethylallyl pyrophosphate (DMAPP) or (E)-4-hydroxy-3-methyl-but-2-enyl diphosphate (HMBPP) to N6-amino group of free or tRNA-bound adenosine phosphates. The reactions are catalyzed by related but distinct enzymes, adenylate dimethylallyltransferase (EC 2.5.1.27) and tRNA dimethylallyltransferase (EC 2.5.1.75), respectively, more frequently termed as isopentenyl transferases Nostoc sp.