In this study, the interaction between gold nanoparticles (AuNPs) and proteins (including lysozyme, trypsin, pepsin, γ-globulin and hemoglobin) was investigated by UV-visible absorption spectroscopy, fluorescence spectroscopy, circular dichroism (CD) spectroscopy and protein activity assay. AuNPs was synthesized using reduction of HAuCl4 with sodium citrate. The formation of AuNPs was confirmed from the characteristic surface plasmon resonance band at 521 nm and transmission electron microscopy revealed the average particle size was about 10 nm. The results reveal that AuNPs can interact with proteins to form a "protein corona (PC)", but the protein concentration required to form a relatively stable PC is not the same. The quenching mechanism of proteins by AuNPs is arisen from static quenching. The binding constants of AuNPs with proteins are in the range from 106 to 1010 L mol-1, and the order is pepsin > γ-globulin > hemoglobin > trypsin > lysozyme at 298 K. Van der Waals forces and hydrogen bonds are the main forces for the lysozyme-AuNPs system. The interaction between trypsin/pepsin/γ-globulin/hemoglobin and AuNPs is mainly by hydrophobic interaction. The addition of AuNPs has an effect on the secondary structure of proteins as confirmed from CD spectra. The change in secondary structure of different proteins is different and seems to have little relation with the binding constant. The activity of lysozyme/trypsin/pepsin decreases with the addition of AuNPs.
Keywords: Fluorescence spectroscopy; Gold nanoparticles; Proteins.
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