Rat GP-3 is a pancreatic lipase with kinetic properties that differ from colipase-dependent pancreatic lipase.
The pancreas contains three homologous proteins, colipase-dependent pancreatic lipase (PL) and two recently described pancreatic lipase-related proteins, PLRP1 and PLRP2. Rat (r) PLRP2 was first identified as a zymogen granule membrane protein, GP-3. Subsequently, we showed that rPLRP2 could cleave fatty acids from triglycerides, but the kinetic properties of rPLRP2 have not been further investigated. To further characterize rPLRP2, we expressed the recombinant enzyme in a baculovirus system, purified the secreted protein, and measured its kinetic properties. rPLRP2 had a broad pH optimum and the curve was similar to that of rPL. At pH 7.5, rPLRP2 cleaved short, medium, and long chain triglycerides by a kinetic mechanism that did not include interfacial activation. The activity against these substrates was not affected by bile salts. In particular, rPLRP2 did not show the bile salt inhibition typical of PL. Although colipase increased rPLRP2 activity in the presence of bile salts, the increase was only 2- to 5-fold compared to the absolute requirement for colipase that rPL had under these conditions. Finally, rPLRP2 could hydrolyze phospholipids, a substrate poorly hydrolyzed by PL. Our characterization of rPLRP2 demonstrates clear differences among the kinetic properties of rPLRP2 and rPL, rPLRP2, and PLRP2 homologues isolated from guinea pig and coypu pancreas. These findings have important implications for the physiological function of rPLRP2.
Jennens
M L
ML
Department of Pediatrics, Washington University School of Medicine, St. Louis Children's Hospital, MO 63110, USA.
Lowe
M E
ME
D000595:Amino Acid Sequence; D000818:Animals; D016367:Baculoviridae / Q000235:genetics*; D001647:Bile Acids and Salts / Q000378:metabolism; D005822:Genetic Vectors; D006863:Hydrogen-Ion Concentration; D007313:Insecta; D007700:Kinetics; D008049:Lipase / Q000096:biosynthesis / Q000235:genetics*; D008969:Molecular Sequence Data; D010179:Pancreas / Q000201:enzymology*; D010740:Phospholipases / Q000096:biosynthesis / Q000235:genetics*; D051381:Rats; D011994:Recombinant Proteins / Q000096:biosynthesis; D017386:Sequence Homology, Amino Acid
D001647:Bile Acids and Salts; D011994:Recombinant Proteins; D010740:Phospholipases; D008049:Lipase; C075988:pancreatic lipase related protein 2