Interfacial interactions in urease/stearic acid and urease/DOPC binary mixed systems at the air-water and the air-solid interfaces

Abstract

In the present study, the protein-lipid interactions between water-soluble metalloenzyme urease (Urs) and insoluble stearic acid (SA) and 1,2-dioleoyl-sn-glycero-3-phosphocholine (DOPC) were investigated at the air–water interface using the Langmuir-Blodgett (LB) technique. Surface pressure (π) - molecular area (A) isotherms were measured for the pure SA and pure DOPC monolayers in the presence of urease in the water subphase. From the (π – A) isotherms of the binary system, it was found that urease strongly interacts with SA and DOPC. The compressibility of the SA monolayer was reduced with the incorporation of urease; a similar trend was also found with DOPC. Moreover, an association of urease with lipid monolayers was found to be surface pressure (π) dependent. The presence of a little hump in the mixed isotherms of Urs-SA at low surface pressure supports this observation. However, the miscibility studies and the calculations of excess Gibb’s free energy indicate an attractive interaction between urease and SA molecules. The considerable morphological variations observed by atomic force microscopy (AFM) in urease-SA deposited films onto hydrophilic and hydrophobic Si(100) surfaces suggest that urease was strongly bound with the surrounding lipid SA on the hydrophilic surfaces but the urease molecules were rigorously denatured on the hydrophobic surfaces. The AFM analysis of Urs-DOPC mixed LB films at room temperature (RT) and elevated temperature suggests substantial denaturation of urease in a hydrophobic environment leading us to speculate the exposure of tryptophan moieties of the urease molecules to the hydrophobic air phase.