Effects of Concentration and Surface Pressure on MBP Interaction with Cholesterol in Langmuir Films

Predicting the mechanism of MBP binding to cholesterol is meaningful in understanding how MBP participate in lateral membrane organization. The interaction of MBP with cholesterol monolayer was investigated at three surface pressures on 10 mM Tris-HCl buffer with the different concentrations of MBP. The results show that π-A isotherms shift to larger molecular area at all pressures. By means of analyzing π-T curves, a surface pressure increase was obtained. Results indicated that the greater the protein concentration in the subphase, the larger the increase of surface pressure. In addition, changes in monolayer surface morphology and domain formation were performed by AFM. These results provide more direct and convincing evidence for the MBP interaction with cholesterol. The MBP-cholesterol interaction suggests a significant concentrations and surface pressure dependence and is probably governed by hydrogen bonds. The date presented could help to understand at least one of the molecular mechanisms through which MBP affects lateral organization of the cholesterol membrane.