Rapid exchange of pancreatic lipase between triacylglycerol droplets.
Two types of experiments were performed to study the reversibility of interfacial adsorption of pancreatic lipase (PL) to fat droplets during lipolysis. Lipolysis was measured in olive oil/gum arabic emulsions containing radiolabeled triolein in the presence of bile salts and lecithin at rate-limiting concentrations of porcine PL (PPL) or human PL (HPL). The lipolysis rate in a labeled emulsion, i.e. release of [(14)C]oleic acid, was immediately reduced by around 50% upon dilution with an equal amount of an unlabeled emulsion. Further, lipolysis was rapidly and completely suppressed when a non-exchanging lipase inhibitor was present in the second emulsion. These results indicate hopping of lipase between emulsion droplets. Alternative explanations were excluded. Hopping of PL between triolein droplets stabilized with gum arabic at supramicellar bile salt concentrations was observed only in the presence, not in the absence, of lecithin. Displacement from a trioctanoin-water interface of active HPL by an inactive mutant (S152G) was studied in the presence of bile salts by measuring HPL distribution between the water phase and the oil-water interface. Colipase was limiting for HPL binding to the oil-water interface (colipase to lipase molar ratio: 0.5) and, thus, for lipolysis. Upon adding S152G, which has the same affinity for colipase, inactive and active HPL were found to compete for binding at the oil-water interface. When equal amounts of HPL and HPL S152G were used, the lipolysis rate dropped to half the maximum rate recorded with HPL alone, suggesting that half the active HPL was rapidly desorbed from the oil-water interface. Therefore, under various conditions, PL does not remain irreversibly adsorbed to the oil-water interface, but can exchange rapidly between oil droplets, via an equilibrium between soluble and lipid-bound PL.
Haiker
Horst
H
F Hoffmann-La Roche, CH-4070 Basel, Switzerland.
Lengsfeld
Hans
H
Hadváry
Paul
P
Carrière
Frédéric
F
D000818:Animals; D002250:Carbon Radioisotopes / Q000378:metabolism; D003089:Colipases / Q000378:metabolism; D006801:Humans; D007783:Lactones / Q000494:pharmacology; D008049:Lipase / Q000037:antagonists & inhibitors / Q000378:metabolism*; D019301:Oleic Acid / Q000378:metabolism; D000069463:Olive Oil; D000077403:Orlistat; D010179:Pancreas / Q000201:enzymology* / Q000378:metabolism; D010938:Plant Oils / Q000378:metabolism; D013552:Swine / Q000378:metabolism; D013997:Time Factors; D014280:Triglycerides / Q000378:metabolism*; D014304:Triolein / Q000378:metabolism
D002250:Carbon Radioisotopes; D003089:Colipases; D007783:Lactones; D000069463:Olive Oil; D010938:Plant Oils; D014280:Triglycerides; D014304:Triolein; D019301:Oleic Acid; D000077403:Orlistat; D008049:Lipase