How colipase fatty acid interactions mediate adsorption of pancreatic lipase to interfaces

Colipase is a cofactor protein which forms a 1:1 complex with pancreatic lipase. This facilitates lipase adsorption to phosphatidylcholine-rich interfaces, presumably as a consequence of the higher affinity of colipase for such interfaces. According to this model, the presence of colipase in an interface should be sufficient to enable lipase adsorption from the aqueous phase. To test this hypothesis, mixed monolayers of colipase, phosphatidylcholine, and fatty acid at the argon-buffer interface were exposed to lipase injected into the stirred aqueous subphase. Spread colipase remained associated with the lipid monolayer in a surface pressure-and lipid composition-dependent manner. For example, with diacylphosphatidyl-choline alone, colipase remained in the lipid monolayer at surface pressures 120 mN/m, but with pure fatty acid this was increased to similar to 40 mN/m. Contrary to the existing paradigm, the presence of colipase in a lipid monolayer was not sufficient to enable the adsorption of lipase to the interface. Fatty acid was also required, and its ability to enhance lipase adsorption over that observed in the absence of colipase was dependent on the fatty acid and colipase mole fractions. These results support the hypothesis that colipase concentrates fatty acids laterally at its periphery and suggest that, together with lipase-colipase interaction, the fatty acid-rich nano-domain surrounding colipase facilitates lipase adsorption in the 'flap-opened' conformation.