Interactions of isoorientin and its Semi-synthetic analogs with human serum albumin

Isoorientin is a C-glycosyl flavone with a wide range of health beneficial effects and inhibits glycogen synthase kinase 3 beta (GSK-3 beta) potentially against Alzheimer's disease. Its semi-synthetic derivatives have greater potency than isoorientin. The present study was aimed to determine the mechanism of interactions of isoorientin and its derivatives with human serum albumin (HSA) using multi-spectroscopic, microscale thermophoresis (MST) and computational studies. Spectra of steady-state fluorescence, UV-Vis, and time-resolved fluorescence indicated that isoorientin and its derivatives quenched the intrinsic fluorescence of HSA through a static quenching process. Isoorientin and its derivatives had a moderate affinity with HSA (K-a 7.7-14.9 x 10(4) M-1). The binding process was accompanied by an exothermic phenomenon, AG degrees of HSA-isoorientin and its derivatives systems were calculated as from -29.51 kJ mol(-1) to -27.87 kJ mol(-1). Displacement experiments with site-specific markers revealed that isoorientin and its derivatives bind to HSA at site II (subdomain IIIA) only. A reduction in the a-helical content of HSA-isoorientin and its derivatives complex was observed, because the conformational changes was structurally perturbed by the hydrophilic groups of the compounds. Further molecular modeling studies confirmed that the binding of isoorientin and its derivatives to the site II via hydrophobic interaction. The MST results confirmed the interactions between HSA and the compounds of interest. The esterase-like assay studies indicated that isoorientin and its derivatives shared the same binding site in HSA, and their induced structural changes of HSA may have been caused by partial unfolding of HSA. This work helps to understand transport, distribution, bioactivity, and design of flavonoid-based GSK-3 beta inhibitors.

Automated summary

The study investigated the interaction mechanism of isoorientin and its derivatives with human serum albumin (HSA) using various spectroscopic techniques, microscale thermophoresis, and computational studies. The results showed that the compounds bind to HSA through a static quenching process and enter site II via hydrophobic interaction. They induce structural changes in HSA.