Synthesis, spectroscopic, physicochemical properties and binding site analysis of 4-(1H-phenanthro[9,10-d]-imidazol-2-yl)-benzaldehyde fluorescent probe for imaging in cell biology: Experimental and theoretical study.
In this study, the 4-(1H-phenanthro[9,10-d]-imidazol-2-yl)-benzaldehyde (PB1) was investigated as a fluorescent dye. For this reason, the spectroscopic properties in different solvents were thoroughly studied. The experimental data were supported by quantum-chemical calculations using density functional theory. Measurements and theoretical calculations showed that PB1 dye is characterized by the non-monotonic solvatochromism, strongly polar charge transfer excited state, large Stokes' shift, high fluorescence quantum yield and high fluorescence lifetime. Simulations using AutoDock presented in this study, showed that after conjugation with Concanavalin A in the active site with LYS116, the PB1 possesses the highest probability of binding affinity. The interaction between the PB1 dye and the Concanavalin A lectin has been investigated by circular dichroism spectroscopy. Conventional fluorescence microscopy imaging of Candida albicans and Yarrowia lipolytica cells, incubated with the PB1-Concanavalin A, was demonstrated. Results show that the PB1 dye is a photostable low molecular weight fluorescent probe, which emits a blue fluorescence. The results of this study have implications for designing PB1-protein conjugate as a valuable alternative to commercial probes designed for cellular labeling in biological and biomedical research. Calculated LogP value together with LogBCF show that PB1-protein conjugate is a valuable alternative to commercial probes designed for cellular labeling in biological and biomedical research.
Nicolaus Copernicus University, Collegium Medicum, Faculty of Pharmacy, Department of Physical Chemistry, Kurpińskiego 5, 85-950 Bydgoszcz, Poland. Electronic address: przemekk@cm.umk.pl.
Przemysław
P
Krawczyk
UTP University of Science and Technology, Faculty of Chemical Technology and Engineering, Seminaryjna 3, 85-326 Bydgoszcz, Poland.
Beata
B
Jędrzejewska
UTP University of Science and Technology, Faculty of Chemical Technology and Engineering, Seminaryjna 3, 85-326 Bydgoszcz, Poland.
Marek
M
Pietrzak
Wroclaw Medical University, Department of Inorganic Chemistry, Faculty of Pharmacy, Borowska 211a, 50-556 Wroclaw, Poland.
Tomasz
T
Janek
D001547:Benzaldehydes / Q000737:chemistry*; D001665:Binding Sites; D002942:Circular Dichroism; D003208:Concanavalin A / Q000737:chemistry; D005456:Fluorescent Dyes / Q000737:chemistry*; D017433:Protein Structure, Secondary
D001547:Benzaldehydes; D005456:Fluorescent Dyes; D003208:Concanavalin A
AutoDock; Conjugation and bioimaging; Density functional theory; Electronic excited state; Fluorochrome; Solvatochromism