Behaviour of protein (BSA)-lipid (DMPA) mixed monolayer on the spreading order of the individual component.

Surface pressure (π) - mean molecular area (A) isotherms of protein (BSA) - lipid (DMPA) mixed films are examined by varying their ratio and altering the spreading order of BSA and DMPA on the water surface to study the protein-lipid interactions and the corresponding structures and patterns at different interfacial conditions. π-A isotherms and compression-decompression isotherm cycles of protein-lipid mixed monolayers below and above of the isoelectric point of BSA (pI ≈ 4.8) are also examined. Below the isoelectric point of BSA (pH ≈ 4.0), i.e., when BSA is weakly hydrophobic and has net positive charge shows low hysteresis irrespective of the spreading order of the molecules. However, at pH ≈ 7.0, i.e., when the overall charge of BSA is negative and is strongly hydrophobic the protein-lipid mixed films display higher hysteresis value. Besides the properties of the isotherms, the surface morphology and secondary conformations of protein inside the mixed films are obtained from X-ray reflectivity, atomic force microscopy (AFM) and Fourier transform infrared (FTIR) spectroscopy respectively after depositing the mixed films on solid substrates. Nearly similar information is obtained after altering the spreading order of BSA and DMPA, which indicates that the spreading of molecules on the water surface is one of the better ways of forming the lipid-protein mixed film at the air-water interface.