TY - JOUR
T1 - A new method for enzymatic preparation of isopentenyladenine-type and trans-zeatin-type cytokinins with radioisotope-labeling
AU - Takei, Kentaro
AU - Dekishima, Yasumasa
AU - Eguchi, Tadashi
AU - Yamaya, Tomoyuki
AU - Sakakibara, Hitoshi
N1 - Funding Information:
Acknowledgment This study was partly supported by Grants-in-Aids for Scientific Research on Priority Areas (number 12142202 to HS) from the Ministry of Education, Culture, Sports, Science, and Technology, Japan.
PY - 2003/6
Y1 - 2003/6
N2 - 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.
AB - 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.
KW - Adenylate isopentenyltransferase
KW - Cytokinin
KW - Isopentenyladenine
KW - Purine-nucleoside phosphorylase
KW - Radioisotope labeling
KW - trans-Zeatin
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U2 - 10.1007/s10265-003-0098-2
DO - 10.1007/s10265-003-0098-2
M3 - Article
C2 - 12728344
AN - SCOPUS:0037695796
VL - 116
SP - 259
EP - 263
JO - Journal of Plant Research
JF - Journal of Plant Research
SN - 0918-9440
IS - 3
ER -