Nanoparticle-protein interactions: Spectroscopic probing of the adsorption of serum albumin to graphene oxide‑gold nanocomplexes surfaces

Graphene oxide‑gold nanocomposites (GO-AuNCPs) are promising candidates in nanomedicine. They will inevitably bind with biomolecules such as serum albumin (SA) in the body while they enter the organism. The interaction between GO-AuNCPs and human serum albumin (HSA)/bovine serum albumin (BSA) were investigated by using multispectroscopic methods, elucidating the binding principles through molecular simulations. The results of ultraviolet-visible (UV-vis) absorption spectroscopy and steady-state fluorescence spectroscopy indicated that GO-AuNCPs interacted with HSA/BSA with different degrees of interaction. The binding of GO-AuNCPs and HSA/BSA was a spontaneous endothermic reaction, and the quenching mechanism is static quenching. The binding constant (K_a) value of BSA binding to GO-AuNCPs at the same temperature was greater than that for HSA, indicating a stronger affinity of BSA for GO-AuNCPs. Molecular simulation revealed that the binding sites of GO-AuNCPs on HSA/BSA were located within the slits of the subdomains IB and IIIA, rather than within any known binding regions. This significant finding was validated by using of site markers phenylbutazone (PB) and flufenamic acid (FA). Synchronous fluorescence spectroscopy, three-dimensional fluorescence spectroscopy, and circular dichroism (CD) spectroscopy showed that the conformation of HSA/BSA was altered upon the addition of GO-AuNCPs, resulting in slight structural changes of tryptophan and tyrosine residues. Although the secondary structure of HSA/BSA was changed, the α-helix remained dominant. The results provide a theoretical and experimental foundation for developing of safe and effective nanomaterials, which is of great theoretical significance.