Design, synthesis and perception of fluorescently labeled isoprenoid cytokinins

0492202 - ÚEB 2019 RIV GB eng J - Journal Article
Kubiasová, K. - Mik, V. - Nisler, Jaroslav - Hönig, Martin - Husičková, A. - Spíchal, L. - Pěkná, Z. - Šamajová, O. - Doležal, Karel - Plíhal, O. - Benková, E. - Strnad, Miroslav - Plíhalová, Lucie
Design, synthesis and perception of fluorescently labeled isoprenoid cytokinins.
Phytochemistry. Roč. 150, JUN (2018), s. 1-11. ISSN 0031-9422. E-ISSN 1873-3700
R&D Projects: GA MŠMT(CZ) LO1204; GA ČR(CZ) GA16-04184S; GA ČR GAP501/10/1450
Institutional support: RVO:61389030
Keywords : barley aleurone protoplasts * endoplasmic-reticulum * nodule organogenesis * histidine kinase * arabidopsis * receptor * binding * probe * ahk4 * identification * 6-[(3-methylbut-2-en-1-yl)amino]purine * N-6-isopentenyladenine * Isoprenoid * Cytokinin * Fluorescent label * Fluorescent probe * Linker * Competitive receptor bioassay * arr5:gus * Live cell confocal microscopy
OECD category: Plant sciences, botany
Impact factor: 2.905, year: 2018

Isoprenoid cytokinins play a number of crucial roles in the regulation of plant growth and development. To study cytokinin receptor properties in plants, we designed and prepared fluorescent derivatives of 6-[(3-methylbut-2-en-1-yl)amino]purine (N-6-isopentenyladenine, iP) with several fluorescent labels attached to the C2 or N9 atom of the purine moiety via a 2-or 6-carbon linker. The fluorescent labels included dansyl (DS), fluorescein (FC), 7-nitrobenzofurazan (NBD), rhodamine B (RhoB), coumarin (Cou), 7-(diethylamino)coumarin (DEAC) and cyanine 5 dye (Cy5). All prepared compounds were screened for affinity for the Arabidopsis thaliana cytokinin receptor (CRE1/AHK4). Although the attachment of the fluorescent labels to iP via the linkers mostly disrupted binding to the receptor, several fluorescent derivatives interacted well. For this reason, three derivatives, two rhodamine B and one 4-chloro-7-nitrobenzofurazan labeled iP were tested for their interaction with CRE1/AHK4 and Zea mays cytokinin receptors in detail. We further showed that the three derivatives were able to activate transcription of cytokinin response regulator ARR5 in Arabidopsis seedlings. The activity of fluorescently labeled cytokinins was compared with corresponding 6-dimethylaminopurine fluorescently labeled negative controls. Selected rhodamine B C2-labeled compounds 17, 18 and 4-chloro-7-nitrobenzofurazan N9-labeled compound 28 and their respective negative controls (19, 20 and 29, respectively) were used for in planta staining experiments in Arabidopsis thaliana cell suspension culture using live cell confocal microscopy.
Permanent Link: http://hdl.handle.net/11104/0285750