Insight into the Lysozyme-Induced Aggregation of Aromatic Amino Acid-Functionalized Gold Nanoparticles: Impact of the Protein Conjugation and Lipid Corona on the Aggregation Phenomena

Abstract

The aggregation and subsequent precipitation of gold nanoparticles (Au NPs) in the presence of protein molecules restrict the usefulness of NPs in biomedical applications. Till now, the influence of different properties of Au NPs (size, surface charge, surface coatings) and proteins (surface charge, chemical modification, folded and unfolded states) and pH and ionic strength of the solution on the aggregation of both Au NPs and proteins has been thoroughly discussed in the literature. However, the underlying different mechanistic pathways of the protein concentration-dependent aggregation of both Au NPs and proteins are poorly understood. The impact of the lipid corona on the protein-induced Au NP aggregation has remained an unresolved issue. In this context, we investigate the interaction of the negatively charged aromatic amino acid (phenylalanine and tyrosine)-functionalized gold nanoparticles (Au-AA NPs) with the positively charged globular protein lysozyme at different protein concentrations and compare the results with those of conventional citrate-functionalized Au NPs (Au-Cit NPs). Next, we conjugate lipids and proteins to Au NPs to impede the aggregation of Au NPs induced by the lysozyme. Our results reveal that the aggregation mechanism of the Au-AA NPs is distinctly different at low and high protein concentrations with the uniqueness of the Au-AA NPs over the Au-Cit NPs. Furthermore, we find that human serum albumin (HSA) protein-conjugated Au-AA and Au-Cit NPs are more effective in preventing the lysozyme-induced Au NP aggregation than bovine serum albumin (BSA)-conjugated Au NPs. For the first time, we also report the significant role of “hard” and “soft” lipid coronas in the aggregation of amino acid (phenylalanine)-functionalized gold nanoparticles in the presence of lysozyme protein. © 2023 American Chemical Society.