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Literature summary extracted from
- Vacuolar and cytosolic cytokinin dehydrogenases of Arabidopsis thaliana: heterologous expression, purification and properties (2010), Phytochemistry, 71, 1970-1978.
Cloned(Commentary)
| EC Number | Cloned (Comment) | Organism |
|---|---|---|
| 1.5.99.12 | expression of cytosolic isozyme AtCKX7 in Escherichia coli and intracellularly in Pichia pastoris | Arabidopsis thaliana |
| 1.5.99.12 | expression of vacuolar His-tagged isozyme AtCKX1 in Pichia pastoris | Arabidopsis thaliana |
| 1.5.99.12 | expression of vacuolar isozyme AtCKX3 in Pichia pastoris | Arabidopsis thaliana |
Localization
| EC Number | Localization | Comment | Organism | GeneOntology No. | Textmining |
|---|---|---|---|---|---|
| 1.5.99.12 | apoplast | secreted isozymes | Arabidopsis thaliana | 48046 | - |
| 1.5.99.12 | cytosol | isozyme AtCKX7 | Arabidopsis thaliana | 5829 | - |
| 1.5.99.12 | vacuole | isozyme AtCKX1 | Arabidopsis thaliana | 5773 | - |
| 1.5.99.12 | vacuole | isozyme AtCKX3 | Arabidopsis thaliana | 5773 | - |
Molecular Weight [Da]
| EC Number | Molecular Weight [Da] | Molecular Weight Maximum [Da] | Comment | Organism |
|---|---|---|---|---|
| 1.5.99.12 | 57057 | - |
x * 60000, SDS-PAGE, x * 57057, mass spectrometry | Arabidopsis thaliana |
| 1.5.99.12 | 58080 | - |
x * 92000, SDS-PAGE, x * 59720, sequence calculation, x * 58080, mass spectrometry | Arabidopsis thaliana |
| 1.5.99.12 | 59720 | - |
x * 92000, SDS-PAGE, x * 59720, sequence calculation, x * 58080, mass spectrometry | Arabidopsis thaliana |
| 1.5.99.12 | 60000 | - |
x * 60000, SDS-PAGE, x * 57057, mass spectrometry | Arabidopsis thaliana |
| 1.5.99.12 | 64290 | - |
x * 97000, SDS-PAGE, x * 65050, sequence calculation, x * 64290, mass spectrometry | Arabidopsis thaliana |
| 1.5.99.12 | 65050 | - |
x * 97000, SDS-PAGE, x * 65050, sequence calculation, x * 64290, mass spectrometry | Arabidopsis thaliana |
| 1.5.99.12 | 92000 | - |
x * 92000, SDS-PAGE, x * 59720, sequence calculation, x * 58080, mass spectrometry | Arabidopsis thaliana |
| 1.5.99.12 | 97000 | - |
x * 97000, SDS-PAGE, x * 65050, sequence calculation, x * 64290, mass spectrometry | Arabidopsis thaliana |
Organism
| EC Number | Organism | UniProt | Comment | Textmining |
|---|---|---|---|---|
| 1.5.99.12 | Arabidopsis thaliana | - |
isozyme AtCKX7 | - |
| 1.5.99.12 | Arabidopsis thaliana | O22213 | isozyme AtCKX1 | - |
| 1.5.99.12 | Arabidopsis thaliana | Q9LTS3 | isozyme AtCKX3 | - |
Purification (Commentary)
| EC Number | Purification (Comment) | Organism |
|---|---|---|
| 1.5.99.12 | 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 |
| 1.5.99.12 | 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)
| EC Number | Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
|---|---|---|---|---|---|---|---|
| 1.5.99.12 | kinetin + 2,3-dimethoxy-5-methyl-1,4-benzoquinone + H2O | - |
Arabidopsis thaliana | ? | - |
? |  |
| 1.5.99.12 | kinetin riboside + 2,3-dimethoxy-5-methyl-1,4-benzoquinone + H2O | - |
Arabidopsis thaliana | ? | - |
? | |
| 1.5.99.12 | 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 | ? | - |
? | |
| 1.5.99.12 | N6-(2-isopentenyl)adenine + 2,3-dimethoxy-5-methyl-1,4-benzoquinone + H2O | - |
Arabidopsis thaliana | adenine + 3-methylbut-2-enal + ? | - |
? | |
| 1.5.99.12 | 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 + ? | - |
? | |
| 1.5.99.12 | 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 + ? | - |
? | |
| 1.5.99.12 | 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 + ? | - |
? | |
| 1.5.99.12 | 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 + ? | - |
? | |
| 1.5.99.12 | N6-(2-isopentenyl)adenine 9-beta-D-riboside + ferricyanide + H2O | - |
Arabidopsis thaliana | adenine 9-beta-D-riboside + 3-methylbut-2-enal + ferrocyanide | - |
? | |
| 1.5.99.12 | N6-(2-isopentenyl)adenine 9-beta-D-riboside + ferricytochrome c + H2O | - |
Arabidopsis thaliana | adenine 9-beta-D-riboside + 3-methylbut-2-enal + ferrocytochrome c | - |
? | |
| 1.5.99.12 | N6-(2-isopentenyl)adenine 9-beta-D-riboside + NAD+ + H2O | - |
Arabidopsis thaliana | adenine 9-beta-D-riboside + 3-methylbut-2-enal + ? | - |
? | |
| 1.5.99.12 | N6-(2-isopentenyl)adenine 9-beta-D-riboside + O2 + H2O | - |
Arabidopsis thaliana | adenine 9-beta-D-riboside + 3-methylbut-2-enal + ? | - |
? | |
| 1.5.99.12 | 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 | - |
? | |
| 1.5.99.12 | 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 + ? | - |
? | |
| 1.5.99.12 | trans-zeatin + 2,3-dimethoxy-5-methyl-1,4-benzoquinone + H2O | - |
Arabidopsis thaliana | ? | - |
? | |
Subunits
| EC Number | Subunits | Comment | Organism |
|---|---|---|---|
| 1.5.99.12 | ? | x * 60000, SDS-PAGE, x * 57057, mass spectrometry | Arabidopsis thaliana |
| 1.5.99.12 | ? | x * 92000, SDS-PAGE, x * 59720, sequence calculation, x * 58080, mass spectrometry | Arabidopsis thaliana |
| 1.5.99.12 | ? | x * 97000, SDS-PAGE, x * 65050, sequence calculation, x * 64290, mass spectrometry | Arabidopsis thaliana |
Synonyms
| EC Number | Synonyms | Comment | Organism |
|---|---|---|---|
| 1.5.99.12 | AtCKX3 | - |
Arabidopsis thaliana |
| 1.5.99.12 | CKX | - |
Arabidopsis thaliana |
Temperature Optimum [°C]
| EC Number | Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
|---|---|---|---|---|
| 1.5.99.12 | 37 | - |
assay at | Arabidopsis thaliana |
pH Optimum
| EC Number | pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
|---|---|---|---|---|
| 1.5.99.12 | 5 | 7.5 | assay at | Arabidopsis thaliana |
Cofactor
| EC Number | Cofactor | Comment | Organism | Structure |
|---|---|---|---|---|
| 1.5.99.12 | flavin | flavoprotein | Arabidopsis thaliana | |
General Information
| EC Number | General Information | Comment | Organism |
|---|---|---|---|
| 1.5.99.12 | evolution | the CKX gene family of Arabidopsis thaliana comprises seven members | Arabidopsis thaliana |
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