The study of caffeine binding to human serum albumin (CROSBI ID 108310)
Prilog u časopisu | izvorni znanstveni rad | međunarodna recenzija
Podaci o odgovornosti
Kriško, Anita ; Kveder, Marina ; Pečar, Slavko ; Pifat, Greta
engleski
The study of caffeine binding to human serum albumin
The binding of caffeine to human serum albumin (HSA) was reinvestigated with the aim of establishing a suitable way of experimental data analysis. We have found that the results obtained previously by other authors using fluorescence spectroscopy are influenced by the non-negligible artefact known as the inner filter effect due to the absorption of caffeine at the excitation wavelength (290 nm). Therefore, we have performed the correction of the obtained data and estimated the binding constants for caffeine binding to HSA. Furthermore, we have applied electron paramagnetic resonance (EPR) spectroscopy using three different positional isomers of spin labeled stearic acid, doxyl stearates, to study the caffeine-HSA interaction. We have found that upon caffeine binding the hypefine splitting decreases for HSA labeled with 5-doxylstearate. This phenomenon together with the observed changes of the spectral amplitudes indicate the increase in mobility of the reporter groups upon caffeine binding, caused by the increase in the local protein dynamics.
HSA; EPR; caffeine; binding
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