Critical role of micelles in pancreatic lipase activation revealed by small angle neutron scattering.
In the duodenum, pancreatic lipase (PL) develops its activity on triglycerides by binding to the bile-emulsified oil droplets in the presence of its protein cofactor pancreatic colipase (PC). The neutron crystal structure of a PC-PL-micelle complex (Hermoso, J., Pignol, D., Penel, S., Roth, M., Chapus, C., and Fontecilla-Camps, J. C. (1997) EMBO J. 16, 5531-5536) has suggested that the stabilization of the enzyme in its active conformation and its adsorption to the emulsified oil droplets are mediated by a preformed lipase-colipase-micelle complex. Here, we correlate the ability of different amphypathic compounds to activate PL, with their association with PC-PL in solution. The method of small angle neutron scattering with D(2)O/H(2)O contrast variation was used to characterize a solution containing PC-PL complex and taurodeoxycholate micelles. The resulting radius of gyration (56 A) and the match point of the solution indicate the formation of a ternary complex that is similar to the one observed in the neutron crystal structure. In addition, we show that either bile salts, lysophospholipids, or nonionic detergents that form micelles with radii of gyration ranging from 13 to 26 A are able to bind to the PC-PL complex, whereas smaller micelles or nonmicellar compounds are not. This further supports the notion of a micelle size-dependent affinity process for lipase activation in vivo.
Pignol
D
D
Laboratoire de Cristallographie et de Cristallogenèse des Protéines, Institut de Biologie Structurale Jean-Pierre Ebel, CEA-CNRS, 41 rue Jules Horowitz, 38027 Grenoble Cedex 1, France.
Ayvazian
L
L
Kerfelec
B
B
Timmins
P
P
Crenon
I
I
Hermoso
J
J
Fontecilla-Camps
J C
JC
Chapus
C
C
D000818:Animals; D001647:Bile Acids and Salts / Q000494:pharmacology; D003089:Colipases / Q000737:chemistry; D003902:Detergents / Q000494:pharmacology; D017666:Deuterium Oxide; D004789:Enzyme Activation; D008049:Lipase / Q000737:chemistry*; D008823:Micelles; D008958:Models, Molecular; D010179:Pancreas / Q000201:enzymology*; D020799:Pancrelipase / Q000737:chemistry; D010261:Paraoxon / Q000494:pharmacology; D010316:Particle Size; D010743:Phospholipids / Q000494:pharmacology; D012542:Scattering, Radiation; D013657:Taurodeoxycholic Acid / Q000737:chemistry
D001647:Bile Acids and Salts; D003089:Colipases; D003902:Detergents; D008823:Micelles; D010743:Phospholipids; D013657:Taurodeoxycholic Acid; D020799:Pancrelipase; D008049:Lipase; D017666:Deuterium Oxide; D010261:Paraoxon