Structure and functional properties of a multimeric protein αA-Crystallin adsorbed on silver nanoparticle surface

Proteins adsorb onto a nanoparticle surface to form a protein-nanoparticle corona which becomes the identity of the nanoparticle in the cellular environment. Conformation of the protein at the interface influences the cellular uptake of the nanoparticle. Hence, interaction of proteins with nanomaterials is of special significance in the field of biotechnology. Adsorption of protein on the nanoparticle surface is a complex process that depends on the dielectric properties and pH of the medium, surface morphology and surface heterogeneity of the nanoparticle, and the quaternary structure of the protein. Thus, interaction of a large multimeric protein with a nanoparticle will be different from that of small oligomeric proteins. In this article we report the conformational and functional properties of a large oligomeric protein αA-Crystallin, a major constituent of the mammalian eye lens, adsorbed onto silver nanoparticle surface. Selective alkylation of the two cysteine residues at the α-Crystallin domain, followed by ITC study showed that these residues play crucial roles in the interaction process. The chaperone function and the refolding capacity of the protein, which is primarily governed by the α-Crystallin domain, are lost to a significant extent when adsorbed onto AgNP surface. The protein in the interface also shows loss of oligomerization that is linked to the biological activity of the protein. Nonetheless, the protein at bio-nano interface shows resistance to urea unfolding process as compared to protein in the solution phase. This might be due to the coordination of AgNP with two cysteine residues of β8 and β9 region of the α-Crystallin domain that imparts extra stability. The compactness in the structure of the adsorbed protein reduces the dynamics of the subunit exchange, which was confirmed by the FRET study. The secondary structure of αA-Crystallin bound to AgNP at substoichiometric ratio remained native-like.