Modulation of excited-state photodynamics of ESIPT probe 1′-hydroxy-2′-acetonaphthone (HAN) on interaction with bovine serum albumin

Abstract

The 1′-hydroxy-2′-acetonaphthone (HAN) is a well-known excited-state intramolecular proton-transfer (ESIPT) dye that has been used as a useful fluorescence probe to understand various homogenous and microheterogenous media. However, interaction of HAN with proteins is still largely less explored. The present study is aimed to investigate the interaction of HAN with the most abundant animal circulatory protein, bovine serum albumin (BSA), using multi-spectroscopic techniques and molecular docking studies. The photophysical properties of HAN undergo significant modulation upon its interaction with BSA protein. The confinement and reduced polarity experienced by bound HAN into BSA binding pocket causes a large fluorescence enhancement. The binding constant for HAN-BSA complex is evaluated to be ∼4.3 × 104 M−1. Time-resolved fluorescence measurements reveal the exclusive presence of only keto (K*) form and twisted keto rotamer (KR*) form of the dye in the excited state inside the BSA binding pocket, though in the ground state the dye exists entirely in its enol (E) form. Results suggest a quantitative ESIPT reaction for HAN even inside the BSA pocket. Time resolved fluorescence anisotropy studies indicate very tight binding of HAN into the BSA pocket such that independent rotational diffusion of the dye is completely ceased. Molecular docking studies provide best docking conformation and involved interaction forces, suggesting the preferred binding of HAN to the site IIA (Sudlow's site I) of the BSA protein. Observed photophysical results provide valuable insights of HAN–BSA binding interaction, revealing for the first time that the intriguing excited-state equilibrium between K* and KR* forms of HAN is largely shifted towards KR* form, as the dye binds to the BSA pocket. To the best of our knowledge such an intriguing modulation in the excited state prototautomeric conversion process has not been reported before for the studied dye in the presence of any other host. Results of the present study can be useful in understanding the pharmacology of structurally similar dyes/drugs, having relevance to drug delivery, pharmaceutical sciences, sensors, and so on.