Effects of colipase and bile salts on the catalytic activity of human pancreatic lipase. A study using the oil drop tensiometer.
Using the oil drop technique, we studied the effects of colipase and bile salts on the rate of hydrolysis of soybean oil by human pancreatic lipase (HPL) as well as on the interfacial binding. Upon continuously recording the decrease in the interfacial tension with time, a 10-15-fold increase in the HPL activity was found to occur in the presence of colipase. The catalytic rate constants of hydrolysis measured at the oil drop surface were found to be of the same order of magnitude as those obtained with monomolecular films spread at the air-water interface. Biotin-labeled HPL (HPL*) was used to determine the amount of adsorbed enzyme using an ELISA test. Less than 1% of the total amount of injected HPL* molecules was found to have adsorbed to the oil-water interface, and no significant effects of colipase on HPL* binding were observed. No significant changes in the hydrolysis rates or the binding of HPL* were detected in the presence of bile salts at concentrations ranging from below their critical micellar concentration (CMC) up to 100 microM. At the oil-water interface, in the absence or presence of bile salts below their CMC, it can be concluded that the colipase is a true lipase cofactor, i.e, it increases the enzyme turnover (approximately 10-15-fold) and does not affect the interfacial lipase adsorption.
Labourdenne
S
S
UPR 9025 de l'IFR1 du CNRS, Laboratoire de Lipolyse Enzymatique, Marseille, France.
Brass
O
O
Ivanova
M
M
Cagna
A
A
Verger
R
R
D000818:Animals; D001647:Bile Acids and Salts / Q000494:pharmacology*; D001710:Biotin; D003089:Colipases / Q000378:metabolism*; D004797:Enzyme-Linked Immunosorbent Assay; D006801:Humans; D007700:Kinetics; D008049:Lipase / Q000378:metabolism*; D008956:Models, Chemical; D009821:Oils; D010179:Pancreas / Q000201:enzymology; D013499:Surface Properties; D013552:Swine; D014867:Water
D001647:Bile Acids and Salts; D003089:Colipases; D009821:Oils; D014867:Water; D001710:Biotin; D008049:Lipase