Systematic elucidation of interactive unfolding and corona formation of bovine serum albumin with cobalt ferrite nanoparticles
Farooq Ahmad, Ying Zhou, Zhaoxing Ling, Qingqing Xiang, Xing Zhou
Nanoparticles (NPs) are extensively being used in modern nano-based therapies and nano-protein formulations. The exposures to these comprehensively used NPs lead to changes in protein structure and functionality, hence raising grave health issues. In this study, we thoroughly investigated the interaction and adsorption of bovine serum albumin (BSA) with CoFe2O4 NPs by circular dichroism (CD), Fourier transform infrared (FTIR), absorption, and fluorescence spectroscopic techniques, scanning electron microscopy (SEM), X-ray diffraction (XRD), vibrating sample magnetometry (VSM), thermogravimetric analysis (TGA) and dynamic light scattering (DLS). The results indicate that CoFe2O4 NPs cause fluorescence quenching in BSA by a static quenching mechanism. The negative values of van't Hoff thermodynamic expressions (ΔH, ΔSand ΔG) corroborate the spontaneity and exothermic nature of static quenching. The major contributors of higher binding affinity of CoFe2O4 NPs with BSA were van der Waals forces and hydrogen bonding. Furthermore, BSA protein corona formation on CoFe2O4 NPs was confirmed by SEM, TGA, DLS and zeta potential studies. TGA, DLS and zeta potential results confirmed the formation of a thick layer of BSA on CoFe2O4 NPs with negative boost in zeta potential. This coating of BSA over CoFe2O4 NPs leads to a decrease in the magnetic saturation value from 50.4 to 46.2 emu, hence the magnetic character of CoFe2O4 NPs. The development of protein corona on CoFe2O4 NPs was further estimated by comparing the steady state fluorescence quenching and theoretical data. In addition, FTIR, UV-CD, and UV-visible spectroscopy and three dimensional fluorescence techniques confirmed that CoFe2O4 NP binding could induce microenvironment perturbations leading to secondary and tertiary conformation changes in BSA. Furthermore, synchronous fluorescence spectroscopy confirmed the significant changes in microenvironment around tryptophan (Trp) residue caused by CoFe2O4 NPs. The denaturing of BSA biochemistry by CoFe2O4 NPs was investigated by assaying esterase activity.
Circular dichroism, Protein folding, Ligand binding, Nanostructures, Secondary structure, Biochemistry, Materials