Activation of horse PLRP2 by bile salts does not require colipase.
Although structurally similar to pancreatic lipase (PL), the key enzyme of intestinal fat digestion, pancreatic lipase-related protein type 2 (PLRP2) differs from PL in certain functional properties. Notably, PLRP2 has a broader substrate specificity than PL, and unlike that of PL, its activity is not restored by colipase in the presence of bile salts. In the studies presented here, the activation mechanism of horse PLRP2 was studied through active site-directed inhibition experiments, and the results demonstrate fundamental differences with that of PL. The opening of the horse PLRP2 flap occurs as soon as bile salt monomers are present, is accelerated in the presence of micelles, and does not require the presence of colipase. Moreover, in contrast to PL, horse PLRP2 is able to directly interact with a bile salt micelle to form an active binary complex, without the micelle being presented by colipase, as evidenced by molecular sieving experiments. These findings, together with the sensitivity of the horse PLRP2 flap to partial proteolysis, are indicative of a higher flexibility of the flap of horse PLRP2 relative to PL. From these results, it can be concluded that PLRP2 can adopt an active conformation in the intestine, which could be important for the further understanding of the physiological role of PLRP2. Finally, this work emphasizes the essential role of colipase in lipase catalysis at the lipid-water interface in the presence of bile.
Jayne
Sandrine
S
Nutrition humaine et lipides, INSERM-U476, 18 Avenue Mozart, 13009 Marseille, France.
Kerfelec
Brigitte
B
Foglizzo
Edith
E
Granon
Simone
S
Hermoso
Juan
J
Chapus
Catherine
C
Crenon
Isabelle
I
D000818:Animals; D001647:Bile Acids and Salts / Q000494:pharmacology*; D003089:Colipases / Q000302:isolation & purification / Q000378:metabolism*; D004789:Enzyme Activation; D005973:Glutamine; D006736:Horses; D007700:Kinetics; D008049:Lipase / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism*; D008239:Lysine; D008958:Models, Molecular; D011487:Protein Conformation; D017433:Protein Structure, Secondary
D001647:Bile Acids and Salts; D003089:Colipases; D005973:Glutamine; D008049:Lipase; C075988:pancreatic lipase related protein 2; D008239:Lysine