Allouche_2007_Biochemistry_46_15188

Colipase is a key element in lipase-catalyzed dietary lipids hydrolysis. Although devoid of enzymatic activity, colipase promotes pancreatic lipase activity in the physiological intestinal conditions by anchoring the enzyme on the surface of lipid droplets. Polarization modulation infrared reflection absorption spectroscopy combined with Brewster angle microscopy studies was performed on colipase alone and in various lipid environments to obtain a global view of both conformation and orientation and to assess lipid perturbations. We clearly show that colipase fully inserts into a dilaurin monolayer and promotes the formation of lipid/protein domains, whereas in a phospholipid environment its insertion is only partial, limited to the polar head group. In a mixed 70% phosphatidylcholine/30% dilaurin environment, colipase adsorbs to but does not penetrate deeply into the film. It triggers the formation of diglyceride domains under which it would form a rather uniform layer. We also clearly demonstrate that colipase adopts a preferred orientation when dilaurin is present at the interface. In contrast, at a neutral phospholipid interface, the infrared spectra suggest an isotropic orientation of colipase which could explain its incapacity to reverse the inhibitory effects of these lipids on the lipase activity.