CE-LIF of Green Fluorescent Fusion Proteins for Investigation of Phosphorylation

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Authors	MICHALCOVÁ Lenka DOPITOVÁ Radka HEJÁTKO Jan RYVOLOVÁ Markéta VÁLKOVÁ Martina PREISLER Jan
Year of publication	2011
Type	Conference abstract
MU Faculty or unit	Central European Institute of Technology
Citation
Attached files	Michalcova_CE-CE_2011.pdf
Description	In this study, CE-LIF with 405 nm laser diode was used to study AHP5 fused with GFP. AHP5-GFP fusion proteins overexpressed in E. coli were used for initial optimization of CE-LIF technique because of lower concentrations of the fusion protein produced by plant. The stability of the AHP5-GFP sample was examined; the complex was found to be unstable with GFP being released. The GFP release was not affected by either a long-term storage at -80 and -20 C or the addition of one of the inhibitors (EDTA, PMSF). These results were confirmed by the western blotting. The phosphorylation of the sample AHP5-GFP in vitro is under investigation. Gel filtration was used to obtain mass fractions for better understanding electrophoregrams. The mass fractions were investigated by CE-LIF and Western blotting-LIF and Western blotting.
Related projects:	Molecular basis of cell and tissue regulations Multidetection platform for microcolumn separation of proteins and peptides CEITEC - Central European Institute of Technology