An improved in vivo deuterium labeling method for measuring the biosynthetic rate of cytokinins

0359292 - ÚEB 2011 RIV CH eng J - Článek v odborném periodiku
Tarkowski, Petr - Floková, K. - Václavíková, Kateřina - Jaworek, P. - Raus, M. - Nordström, A. - Novák, Ondřej - Doležal, Karel - Šebela, M. - Frébortová, Jitka
An improved in vivo deuterium labeling method for measuring the biosynthetic rate of cytokinins.
Molecules. Roč. 15, č. 12 (2010), s. 9214-9229. E-ISSN 1420-3049
Grant CEP: GA ČR(CZ) GA522/08/0920; GA MŠMT ED0017/01/01; GA ČR GA301/08/1649
Výzkumný záměr: CEZ:AV0Z50380511
Klíčová slova: cytokinin * deuterium labelling * biosynthetic rate
Kód oboru RIV: CE - Biochemie
Impakt faktor: 1.988, rok: 2010

A set of 11 relevant isoprenoid cytokinins, including zeatin isomers, was separated by ultra performance liquid chromatography in less than 6 min. The iP-type cytokinins were observed to give rise to a previously-unknown fragment at m/z 69; we suggest that the diagnostic (204-69) transition can be used to monitor the biosynthetic rate of isopentenyladenine. Furthermore, we found that by treating the cytokinin nucleotides with alkaline phosphatase prior to analysis, the sensitivity of the detection process could be increased. In addition, derivatization (propionylation) improved the ESIMS response by increasing the analytes' hydrophobicity. Indeed, the ESI-MS response of propionylated isopentenyladenosine was about 34% higher than that of its underivatized counterpart. Moreover, the response of the derivatized zeatin ribosides was about 75% higher than that of underivatized zeatin ribosides. Finally, we created a web-based calculator (IZOTOP) that facilitates MS/MS data processing.
Trvalý link: http://hdl.handle.net/11104/0197101