NDLI: An Improved in Vivo Deuterium Labeling Method for Measuring the Biosynthetic Rate of Cytokinins

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An Improved in Vivo Deuterium Labeling Method for Measuring the Biosynthetic Rate of Cytokinins

Content Provider	MDPI
Author	Tarkowski, Petr Floková, Kristýna Václavíková, Kateřina Jaworek, Pavel Raus, Martin Nordström, Anders Novák, Ondřej Doležal, Karel Šebela, Marek Frébortová, Jitka
Abstract	An improved method for determining the relative biosynthetic rate of isoprenoid cytokinins has been developed. 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 ESI-MS 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 and offer it freely to the research community.
File Size	259072
Ending Page	9229
Page Count	16
Starting Page	9214
File Format	PDF
e-ISSN	14203049
DOI	10.3390/molecules15129214
Journal	Molecules
Issue Number	12
Volume Number	15
Language	English
Publisher Date	2010-12-15
Access Restriction	Open
Subject Keyword	cytokinin deuterium labelling biosynthetic rate UPLC MS
Content Type	Text
Resource Type	Article

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