A new method for enzymatic preparation of isopentenyladenine-type and trans-zeatin-type cytokinins with radioisotope-labeling

Kentaro Takei, Yasumasa Dekishima, Tadashi Eguchi, Tomoyuki Yamaya, Hitoshi Sakakibara

Research output: Contribution to journalArticlepeer-review

14 Citations (Scopus)

Abstract

We describe a new enzymatic reaction method for the preparation of the radioisotope-labeled cytokinins isopentenyladenine (iP), trans-zeatin (tZ), and their ribosides. The method is based on the three enzyme activities of an adenylate isopentenyltransferase (IPT; EC 2.5.1.27) from Arabidopsis thaliana, an alkaline phosphatase (EC 3.1.3.1) from calf intestine, and a purine-nucleoside phosphorylase (EC 2.4.2.1) from Escherichia coli. The A. thaliana IPT, AtIPT7, utilized both dimethylallyldiphosphate and 4-hydroxy-3-methyl-2-(E)-butenyl diphosphate as isoprenoid donors. The dual specificity of the substrates enabled us to produce iP-type and tZ-type cytokinins separately in the same system simply by switching the substrates. Our method affords a much higher yield of the labeled products than the chemical reaction methods previously used. These labeled compounds will be useful tools for cytokinin research, such as receptor-ligand assays and cell metabolism studies.

Original languageEnglish
Pages (from-to)259-263
Number of pages5
JournalJournal of Plant Research
Volume116
Issue number3
DOIs
Publication statusPublished - 2003 Jun
Externally publishedYes

Keywords

  • Adenylate isopentenyltransferase
  • Cytokinin
  • Isopentenyladenine
  • Purine-nucleoside phosphorylase
  • Radioisotope labeling
  • trans-Zeatin

ASJC Scopus subject areas

  • Plant Science

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