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Literature summary for 1.5.99.12 extracted from

  • Kowalska, M.; Galuszka, P.; Frebortova, J.; Sebela, M.; Beres, T.; Hluska, T.; Smehilova, M.; Bilyeu, K.D.; Frebort, I.
    Vacuolar and cytosolic cytokinin dehydrogenases of Arabidopsis thaliana: heterologous expression, purification and properties (2010), Phytochemistry, 71, 1970-1978.
    View publication on PubMed

Cloned(Commentary)

Cloned (Comment) Organism
expression of cytosolic isozyme AtCKX7 in Escherichia coli and intracellularly in Pichia pastoris Arabidopsis thaliana
expression of vacuolar His-tagged isozyme AtCKX1 in Pichia pastoris Arabidopsis thaliana
expression of vacuolar isozyme AtCKX3 in Pichia pastoris Arabidopsis thaliana

Localization

Localization Comment Organism GeneOntology No. Textmining
apoplast secreted isozymes Arabidopsis thaliana 48046
-
cytosol isozyme AtCKX7 Arabidopsis thaliana 5829
-
vacuole isozyme AtCKX1 Arabidopsis thaliana 5773
-
vacuole isozyme AtCKX3 Arabidopsis thaliana 5773
-

Molecular Weight [Da]

Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
57057
-
x * 60000, SDS-PAGE, x * 57057, mass spectrometry Arabidopsis thaliana
58080
-
x * 92000, SDS-PAGE, x * 59720, sequence calculation, x * 58080, mass spectrometry Arabidopsis thaliana
59720
-
x * 92000, SDS-PAGE, x * 59720, sequence calculation, x * 58080, mass spectrometry Arabidopsis thaliana
60000
-
x * 60000, SDS-PAGE, x * 57057, mass spectrometry Arabidopsis thaliana
64290
-
x * 97000, SDS-PAGE, x * 65050, sequence calculation, x * 64290, mass spectrometry Arabidopsis thaliana
65050
-
x * 97000, SDS-PAGE, x * 65050, sequence calculation, x * 64290, mass spectrometry Arabidopsis thaliana
92000
-
x * 92000, SDS-PAGE, x * 59720, sequence calculation, x * 58080, mass spectrometry Arabidopsis thaliana
97000
-
x * 97000, SDS-PAGE, x * 65050, sequence calculation, x * 64290, mass spectrometry Arabidopsis thaliana

Organism

Organism UniProt Comment Textmining
Arabidopsis thaliana
-
isozyme AtCKX7
-
Arabidopsis thaliana O22213 isozyme AtCKX1
-
Arabidopsis thaliana Q9LTS3 isozyme AtCKX3
-

Purification (Commentary)

Purification (Comment) Organism
recombinant vacuolar His-tagged isozyme AtCKX1 41.7fold from Pichia pastoris by ultrafiltration, hydrophobic interaction and hydroxyapatite chromatography, followed by nickel affinity chromatography and another step of hydrophobic interaction chromatography Arabidopsis thaliana
recombinant vacuolar isozyme AtCKX3 18fold from Pichia pastoris by ultrafiltration, hydroxyapatite chromatography, nickel affinity chromatography, and anion exchange chromatography and another step of hydrophobic interaction chromatography Arabidopsis thaliana

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
kinetin + 2,3-dimethoxy-5-methyl-1,4-benzoquinone + H2O
-
Arabidopsis thaliana ?
-
?
kinetin riboside + 2,3-dimethoxy-5-methyl-1,4-benzoquinone + H2O
-
Arabidopsis thaliana ?
-
?
additional information acceptor and substrate specificity, overview. The enzyme catalyzes the irreversible cleavage of N6-side chains from cytokinins. CKXs also show low cytokinin oxidase activity, but molecular oxygen is a comparatively poor electron acceptor. Vacuolar AtCKX enzymes in certain conditions degrade N6-(2-isopentenyl)adenine di- and triphosphates two to 5times more effectively than its monophosphate Arabidopsis thaliana ?
-
?
N6-(2-isopentenyl)adenine + 2,3-dimethoxy-5-methyl-1,4-benzoquinone + H2O
-
Arabidopsis thaliana adenine + 3-methylbut-2-enal + ?
-
?
N6-(2-isopentenyl)adenine 9-beta-D-glucoside + 3',5'-dimethoxy-4'-hydroxyacetophenone + H2O i.e. iP9G, best substrate for isozyme AtCKX1 at pH 5.0, 3',5'-dimethoxy-4-hydroxyacetophenone is the best acceptor Arabidopsis thaliana adenine 9-beta-D-glucoside + 3-methylbut-2-enal + ?
-
?
N6-(2-isopentenyl)adenine 9-beta-D-riboside + 2,3,5-triphenyl-tetrazolium chloride + H2O
-
Arabidopsis thaliana adenine 9-beta-D-riboside + 3-methylbut-2-enal + ?
-
?
N6-(2-isopentenyl)adenine 9-beta-D-riboside + 2,3-dimethoxy-5-methyl-1,4-benzoquinone + H2O
-
Arabidopsis thaliana adenine 9-beta-D-riboside + 3-methylbut-2-enal + ?
-
?
N6-(2-isopentenyl)adenine 9-beta-D-riboside + 3',5'-dimethoxy-4'-hydroxyacetophenone + H2O
-
Arabidopsis thaliana adenine 9-beta-D-riboside + 3-methylbut-2-enal + ?
-
?
N6-(2-isopentenyl)adenine 9-beta-D-riboside + ferricyanide + H2O
-
Arabidopsis thaliana adenine 9-beta-D-riboside + 3-methylbut-2-enal + ferrocyanide
-
?
N6-(2-isopentenyl)adenine 9-beta-D-riboside + ferricytochrome c + H2O
-
Arabidopsis thaliana adenine 9-beta-D-riboside + 3-methylbut-2-enal + ferrocytochrome c
-
?
N6-(2-isopentenyl)adenine 9-beta-D-riboside + NAD+ + H2O
-
Arabidopsis thaliana adenine 9-beta-D-riboside + 3-methylbut-2-enal + ?
-
?
N6-(2-isopentenyl)adenine 9-beta-D-riboside + O2 + H2O
-
Arabidopsis thaliana adenine 9-beta-D-riboside + 3-methylbut-2-enal + ?
-
?
N6-(2-isopentenyl)adenine 9-beta-D-riboside + oxidized 2,6-dichlorophenolindophenol + H2O
-
Arabidopsis thaliana adenine 9-beta-D-riboside + 3-methylbut-2-enal + reduced 2,6-dichlorophenolindophenol
-
?
N6-(2-isopentenyl)adenine 9-beta-D-riboside 5'-phosphate + 2,3-dimethoxy-5-methyl-1,4-benzoquinone + H2O
-
Arabidopsis thaliana adenine 9-beta-D-riboside 5'-phosphate + 3-methylbut-2-enal + ?
-
?
trans-zeatin + 2,3-dimethoxy-5-methyl-1,4-benzoquinone + H2O
-
Arabidopsis thaliana ?
-
?

Subunits

Subunits Comment Organism
? x * 60000, SDS-PAGE, x * 57057, mass spectrometry Arabidopsis thaliana
? x * 92000, SDS-PAGE, x * 59720, sequence calculation, x * 58080, mass spectrometry Arabidopsis thaliana
? x * 97000, SDS-PAGE, x * 65050, sequence calculation, x * 64290, mass spectrometry Arabidopsis thaliana

Synonyms

Synonyms Comment Organism
AtCKX3
-
Arabidopsis thaliana
CKX
-
Arabidopsis thaliana

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
37
-
assay at Arabidopsis thaliana

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
5 7.5 assay at Arabidopsis thaliana

Cofactor

Cofactor Comment Organism Structure
flavin flavoprotein Arabidopsis thaliana

General Information

General Information Comment Organism
evolution the CKX gene family of Arabidopsis thaliana comprises seven members Arabidopsis thaliana