Effect of Acetonitrile on the Conformation of Bovine Serum Albumin

• Published in: ACS omega 2024-12, Vol.9 (48), p.47680-47689
• Main Authors: Kaumbekova, Samal, Sugita, Masatake, Sakaguchi, Naoya, Takahashi, Yuta, Sadakane, Akira, Umezawa, Masakazu
• Format: Article
• Language: English
• ISSN: 2470-1343; 2470-1343
• DOI: 10.1021/acsomega.4c07274

The use of organic solvents in drug delivery systems (DDSs) either to produce albumin nanoparticles or to manipulate the binding of target molecules to albumin, a promising nanocarrier material, presents challenges due to the conformational changes induced in the protein. In this study, we investigated the alterations in the conformation of bovine serum albumin (BSA) caused by acetonitrile (ACN) in aqueous solution by using a combination of spectroscopic analysis and molecular dynamics (MD) simulations. Ultraviolet (UV) absorption, fluorescence, and infrared (IR) absorption spectroscopy were used to analyze the BSA conformation in the solutions containing 0–60 vol % ACN. Additionally, MD simulations were conducted to elucidate the interactions between BSA and solvent components, focusing on the structural changes in the hydrophobic pocket with Trp residues of the albumin. Increasing the ACN concentration leads to significant changes in the BSA conformation, as evidenced by shifts in UV fluorescence wavelength, decreased intensity, and alterations in IR absorption bands. Furthermore, the formation of protein aggregates was observed at high ACN concentration (30 vol % ACN), shown by increased hydrodynamic diameter distribution. MD simulations further demonstrate that the presence of ACN molecules near the hydrophobic pocket with the Trp-213 residue increases the fluctuations in the positions of amino acids observed near the hydrophobic pocket with Trp-213. Moreover, the intrusion of water molecules into the hydrophobic pocket under 60% ACN conditions with highly decreased solvent polarity was correlated with the changes in the BSA secondary structure. These findings enhance our understanding of how solvent polarity affects the albumin conformation, which is crucial for optimizing albumin-based DDS applications.