Information on EC 2.5.1.27 - adenylate dimethylallyltransferase

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The expected taxonomic range for this enzyme is: Bacteria, Eukaryota

EC NUMBER
COMMENTARY hide
2.5.1.27
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RECOMMENDED NAME
GeneOntology No.
adenylate dimethylallyltransferase
REACTION
REACTION DIAGRAM
COMMENTARY hide
ORGANISM
UNIPROT
LITERATURE
dimethylallyl diphosphate + AMP = diphosphate + N6-(dimethylallyl)adenosine 5'-phosphate
show the reaction diagram
REACTION TYPE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
alkenyl group transfer
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-
-
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PATHWAY
BRENDA Link
KEGG Link
MetaCyc Link
Biosynthesis of secondary metabolites
-
-
trans-zeatin biosynthesis
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Zeatin biosynthesis
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SYSTEMATIC NAME
IUBMB Comments
dimethylallyl-diphosphate:AMP dimethylallyltransferase
Involved in the biosynthesis of cytokinins in plants. Some isoforms from the plant Arabidopsis thaliana are specific for AMP while others also have the activity of EC 2.5.1.112, adenylate dimethylallyltransferase (ADP/ATP-dependent).
CAS REGISTRY NUMBER
COMMENTARY hide
72840-95-0
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ORGANISM
COMMENTARY hide
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
-
-
-
Manually annotated by BRENDA team
-
UniProt
Manually annotated by BRENDA team
Pineapple sweet orange
-
-
Manually annotated by BRENDA team
Carrizo citrange
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-
Manually annotated by BRENDA team
soybean
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-
Manually annotated by BRENDA team
gene SAG12
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-
Manually annotated by BRENDA team
mulberry
SwissProt
Manually annotated by BRENDA team
isoform isopentenyl transferase 2, IPT2
UniProt
Manually annotated by BRENDA team
GENERAL INFORMATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
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
metabolism
physiological function
-
the isopentenyltransferase reaction is the key rate-limiting step in cytokinin biosynthesis that transfers the isopentenyl group from dimethylallyl diphosphate to the N6-amino group of adenine
additional information
SUBSTRATE
PRODUCT                       
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
(E)-4-hydroxy-3-methyl-but-2-enyl diphosphate + AMP
diphosphate + trans-zeatin-5'-monophosphate
show the reaction diagram
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 dphosphate
-
-
?
1-hydroxy-2-methyl-2-(E)-butenyl 4-diphosphate + AMP
diphosphate + N6-(4-hydroxy-3-methyl-2-(E)-butyl)adenosine 5'-phosphate
show the reaction diagram
4-hydroxy-3-methyl-2-(E)-butenyl diphosphate + ADP
diphosphate + N6-(4-hydroxy-3-methyl-2-(E)-butenyl)ADP
show the reaction diagram
4-hydroxy-3-methyl-2-(E)-butenyl diphosphate is a very poor substrate
-
-
?
4-hydroxy-3-methyl-2-(E)-butenyl diphosphate + AMP
diphosphate + N6-(4-hydroxy-3-methyl-2-(E)-butyl)adenosine 5'-phosphate
show the reaction diagram
-
-
-
-
?
4-hydroxy-3-methyl-2-(E)-butenyl diphosphate + ATP
diphosphate + N6-(4-hydroxy-3-methyl-2-(E)-butenyl)ATP
show the reaction diagram
4-hydroxy-3-methyl-2-(E)-butenyl diphosphate is a very poor substrate
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-
?
dimethylallyl diphosphate + ADP
?
show the reaction diagram
dimethylallyl diphosphate + ADP
dimethylallyl-ADP + diphosphate
show the reaction diagram
the enzyme preferrs ADP and ATP rather than AMP
-
-
?
dimethylallyl diphosphate + ADP
diphosphate + N-(dimethylallyl)adenosine 5'-diphosphate
show the reaction diagram
ADP and ATP are preferred over AMP
-
-
?
dimethylallyl diphosphate + ADP
diphosphate + N6-(dimethylallyl)adenosine 5'-diphosphate
show the reaction diagram
dimethylallyl diphosphate + ADP
isopentenyl-ADP + diphosphate
show the reaction diagram
-
-
-
-
?
dimethylallyl diphosphate + AMP
diphosphate + N-(dimethylallyl)adenosine 5'-phosphate
show the reaction diagram
dimethylallyl diphosphate + AMP
diphosphate + N6-(dimethylallyl)adenosine 5'-phosphate
show the reaction diagram
dimethylallyl diphosphate + ATP
?
show the reaction diagram
dimethylallyl diphosphate + ATP
dimethylallyl-ATP + diphosphate
show the reaction diagram
the enzyme preferrs ADP and ATP rather than AMP
-
-
?
dimethylallyl diphosphate + ATP
diphosphate + N-(dimethylallyl)adenosine 5'-triphosphate
show the reaction diagram
ADP and ATP are preferred over AMP
-
-
?
dimethylallyl diphosphate + ATP
diphosphate + N6-(dimethylallyl)adenosine 5'-triphosphate
show the reaction diagram
dimethylallyl diphosphate + ATP
diphosphate + N6-(dimethylallyl)adenosine triphosphate
show the reaction diagram
-
-
-
-
?
dimethylallyl diphosphate + ATP
N6-(dimethylallyl) adenosine 5'-triphosphate + diphosphate
show the reaction diagram
-
-
-
-
ir
dimethylallyl diphosphate + CDP
?
show the reaction diagram
-
-
-
ir
dimethylallyl diphosphate + dADP
diphosphate + N6-(dimethylallyl)dADP
show the reaction diagram
-
-
-
ir
dimethylallyl diphosphate + dATP
diphosphate + N6-(dimethylallyl)dATP
show the reaction diagram
-
-
-
ir
dimethylallyl diphosphate + GDP
?
show the reaction diagram
GDP is a poor substrate
-
-
ir
dimethylallyl diphosphate + P1,P3-bis(5'-adenosyl)triphosphate
?
show the reaction diagram
-
-
-
-
?
dimethylallyl diphosphate + P1,P4-bis(5'-adenosyl)tetraphosphate
?
show the reaction diagram
-
-
-
-
?
dimethylallyl diphosphate + P1,P5-bis(5'-adenosyl)pentaphosphate
?
show the reaction diagram
-
-
-
-
?
dimethylallyl diphosphate + P1,P6-bis(5'-adenosyl)hexaphosphate
?
show the reaction diagram
-
-
-
-
?
geranyl diphosphate + ADP
diphosphate + N6-geranyl-ADP
show the reaction diagram
geranyl diphosphate is a very poor substrate
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-
?
isopentenyl diphosphate + ADP
diphosphate + N6-isopentyl-ADP
show the reaction diagram
isopentyl diphosphate is a very poor substrate
-
-
?
additional information
?
-
NATURAL SUBSTRATES
NATURAL PRODUCTS
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
REVERSIBILITY
r=reversible
ir=irreversible
?=not specified
1-hydroxy-2-methyl-2-(E)-butenyl 4-diphosphate + AMP
diphosphate + N6-(4-hydroxy-3-methyl-2-(E)-butyl)adenosine 5'-phosphate
show the reaction diagram
-
1-hydroxy-2-methyl-2-(E)-butenyl 4-diphosphate is an intermediate in the methylerythritol phosphate pathway. Agrobacterium tumefaciens modifies cytokinin biosynthesis by sending a key enzyme into plastids of the host plant to promote tumorigenesis
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-
?
dimethylallyl diphosphate + AMP
diphosphate + N-(dimethylallyl)adenosine 5'-phosphate
show the reaction diagram
-
-
-
-
ir
dimethylallyl diphosphate + AMP
diphosphate + N6-(dimethylallyl)adenosine 5'-phosphate
show the reaction diagram
dimethylallyl diphosphate + ATP
N6-(dimethylallyl) adenosine 5'-triphosphate + diphosphate
show the reaction diagram
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-
-
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ir
additional information
?
-
COFACTOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
METALS and IONS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
MgCl2
dependent on presence of divalent cation, addition of 2 mM MnCl2 is 1.3 times more effective than that of 10 mM MgCl2
MnCl2
dependent on presence of divalent cation, addition of 2 mM MnCl2 is 1.3 times more effective than that of 10 mM MgCl2
Zn2+
two Zn2+ ions are present in the crystal. One is coordinated with the hydroxyl group of Thr-15. The other is located near AMP and is coordinated to the side chains of Asp-173, Glu-213, and His-214
INHIBITORS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.00023 - 0.0805
1-hydroxy-2-methyl-2-(E)-butenyl 4-diphosphate
0.0147 - 0.0682
ADP
0.000086 - 0.759
AMP
0.0051 - 0.018
ATP
0.000008 - 0.05
dimethylallyl diphosphate
additional information
additional information
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TURNOVER NUMBER [1/s]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.00023 - 0.025
1-hydroxy-2-methyl-2-(E)-butenyl 4-diphosphate
0.0235 - 0.024
ADP
0.00828
AMP
Humulus lupulus
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pH 8.0, 25C
0.022
ATP
Humulus lupulus
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pH 8.0, 25C
0.002 - 0.09
dimethylallyl diphosphate
SPECIFIC ACTIVITY [µmol/min/mg]
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
0.01122
purified recombinant enzyme, pH 7.5, 25C
additional information
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assay principle
pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
5 - 7
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assay at
6 - 9
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either in Tris-HCl or potassium-phosphate buffer
TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
37
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assay at
SOURCE TISSUE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
SOURCE
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cytokinin-autotrophic
Manually annotated by BRENDA team
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crown gall tumor line 15055/01
Manually annotated by BRENDA team
at 8-10 days after pollination the endosperm and especially the basal transfer cell layer is a major site of IPT2 expression, this expression persists in the basal transfer cell layer and the developing embryo into later kernel development stages
Manually annotated by BRENDA team
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fruit abscission zone
Manually annotated by BRENDA team
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upper part of younger inflorescences
Manually annotated by BRENDA team
strong expression of isoform IPT2 in developing kernels. At 8-10 days after pollination the endosperm and especially the basal transfer cell layer is a major site of IPT2 expression, this expression persists in the basal transfer cell layer and the developing embryo into later kernel development stages
Manually annotated by BRENDA team
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AtIPT3, upregulated within 1 h after an application of nitrate zo mineral-strved Arabidopsis plants
Manually annotated by BRENDA team
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immature, AtIPT1; immature, AtIPT4
Manually annotated by BRENDA team
very low level of isoform IPT2 protein
Manually annotated by BRENDA team
LOCALIZATION
ORGANISM
UNIPROT
COMMENTARY hide
GeneOntology No.
LITERATURE
SOURCE
-
farnesylated protein
Manually annotated by BRENDA team
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farnesylated protein
Manually annotated by BRENDA team
PDB
SCOP
CATH
ORGANISM
UNIPROT
Agrobacterium fabrum (strain C58 / ATCC 33970)
Agrobacterium fabrum (strain C58 / ATCC 33970)
Agrobacterium fabrum (strain C58 / ATCC 33970)
Agrobacterium fabrum (strain C58 / ATCC 33970)
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
35810
calculated from sequence of cDNA
37000
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x * 37000, SDS-PAGE
40000
x * 40000, recombinant His6-tagged BoAIPT1, SDS-PAGE
SUBUNITS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
POSTTRANSLATIONAL MODIFICATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
alkylation
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farnesylation
Crystallization/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
isoform Tzs in complex with AMP and dimethylallyl S-thiodiphosphate. The carbon-nitrogen-based prenylation proceeds by the SN2-reaction mechanism
The crystal structure of the AIPT-ATP complex from Humulus lupulus is similar to the previous structures of Agrobacterium AIPT and yeast tRNA-IPT. The enzyme is structurally homologous to the NTP-binding kinase family of proteins but forms a solvent-accessible channel that binds to the donor substrate dimethylallyl diphosphate, which is directed toward the acceptor substrate ATP/ADP.
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TEMPERATURE STABILITY
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
additional information
-
low thermal stability
STORAGE STABILITY
ORGANISM
UNIPROT
LITERATURE
-70C, about 90% loss of activity after 3 weeks
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Purification/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
Ni-chelate affinity column chromatography
Ni-NTA column chromatography; recombinant His-tagged enzyme from Escherichia coli strain BL21 (DE3) STAR by anion exchange and nickel affinity chromatography to apparent homogeneity
recombinant enzyme
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recombinant His6-tagged BoAIPT1 from Escherichia coli strain BL21 (DE3) by nickel affinity chromatography and ultrafiltration
Cloned/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
cDNA encoding BoAIPT1, DNA and amino acid sequence determination and analysis, sequence comparison, functional expression of N-terminally His6-tagged BoAIPT1 in Escherichia coli strain BL21 (DE3), optimum protein expression is obtained by incubating the IPTG-induced culture at 18C for 18 hr, transgenic expression in bamboo plants, real-time PCR expression analysis
cloning from leaves, expression of IPT in transgenic peanut plants by transfection using Agrobacterium tumefaciens
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construct consisting of the 35S promoter fused to the first intron of the maize alcohol dehydrogenase gene, fused to the isoform IPT2 coding sequence and a terminator. Ectopic expression in Arabidopsis thaliana
expressed as fusions protein in Escherichia coi and Saccharomyces cerevisiae strains CTY716 and SYY705
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expressed in Arabidopsis thaliana
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expressed in Arabidopsis thaliana (root, leaf and flower)
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expressed in Escherichia coli
expressed in Escherichia coli Top10 cells; 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
expressed in Escherichia coli; expressed in Escherichia coli
expression in Escherichia coli
fusion with a seeds-pecific lectin promoter from Glycine max and expression in Nicotiana tabacum
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gene ipt, expression in transgenic Oryza sativa plants via Agrobacterium tumefaciens strain LBA4404 transfection using the glutenin HMW promoter
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gene SAG12, overexpression in transgenic cassava plants, transgene integration patterns in SAG12-IPT transgenic lines, overview
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overexpressed as a N-terminal hexahistidine tag protein in Escherichia coli BL21 (DE3)
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EXPRESSION
ORGANISM
UNIPROT
LITERATURE
the SAG12 gene is senescence-induced in leaves
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ENGINEERING
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
D173G
decrease in activity with substrate 1-hydroxy-2-methyl-2-(E)-butenyl 4-diphosphate. Specificity ratio for substrate dimethylallyl diphosphate over 1-hydroxy-2-methyl-2-(E)-butenyl 4-diphosphate is 208 compared with 2.3 for wild-type
E210N
slight decrease in activity with substrate 1-hydroxy-2-methyl-2-(E)-butenyl 4-diphosphate. Specificity ratio for substrate dimethylallyl diphosphate over 1-hydroxy-2-methyl-2-(E)-butenyl 4-diphosphate is 15 compared with 2.3 for wild-type
E213Q
slight decrease in activity with substrate 1-hydroxy-2-methyl-2-(E)-butenyl 4-diphosphate. Specificity ratio for substrate dimethylallyl diphosphate over 1-hydroxy-2-methyl-2-(E)-butenyl 4-diphosphate is 5.1 compared with 2.3 for wild-type
H214L
decrease in activity with substrate 1-hydroxy-2-methyl-2-(E)-butenyl 4-diphosphate. Specificity ratio for substrate dimethylallyl diphosphate over 1-hydroxy-2-methyl-2-(E)-butenyl 4-diphosphate is 1163 compared with 2.3 for wild-type
Y211T
slight decrease in activity with substrate 1-hydroxy-2-methyl-2-(E)-butenyl 4-diphosphate. Specificity ratio for substrate dimethylallyl diphosphate over 1-hydroxy-2-methyl-2-(E)-butenyl 4-diphosphate is 9 compared with 2.3 for wild-type
D49A
almost complete loss of activity compared to the wild type enzyme
F120A
almost complete loss of activity compared to the wild type enzyme
F157A
almost complete loss of activity compared to the wild type enzyme
F93A
almost complete loss of activity compared to the wild type enzyme
Q255A
almost complete loss of activity compared to the wild type enzyme
W159A
almost complete loss of activity compared to the wild type enzyme
Y153A
almost complete loss of activity compared to the wild type enzyme
Y170A
retains 84% activity compared to the wild type enzyme
Y217A
almost complete loss of activity compared to the wild type enzyme
Y54A
almost complete loss of activity compared to the wild type enzyme
additional information
APPLICATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
synthesis
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method for enzymatic preparation of isopentenyladenine-type and trans-zeatin-type cytokinins with radioisotope-labeling. The method is based on EC 2.5.1.27 from Arabidopsis thaliana, EC 3.1.3.1 from calf intestine and EC 2.4.2.1 from Escherichia coli