Characterization of pancreatic lipase-related protein 2 isolated from human pancreatic juice
Autor: | Josiane De Caro, Philippe Grandval, Hubert Halimi, Francine Ferrato, Alain De Caro, Frédéric Carrière, Barbara Sias |
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Rok vydání: | 2004 |
Předmět: |
Chromatography
biology Biophysics Galactolipids Lipase Colipase Biochemistry Antibodies Analytical Chemistry Bile Acids and Salts Affinity chromatography Pancreatic Juice Polyclonal antibodies Galactolipase Phospholipases Pancreatic juice biology.protein Humans Colipases Molecular Biology Carboxylic Ester Hydrolases Galactolipase activity |
Zdroj: | Biochimica et biophysica acta. 1701(1-2) |
ISSN: | 0006-3002 |
Popis: | Human pancreatic lipase-related protein 2 (HPLRP2) was identified for the first time in pancreatic juice using specific anti-peptide antibodies and purified to homogeneity. Antibodies were raised in the rabbit using a synthetic peptide from the HPLRP2 protein sequence deduced from cDNA. Western blotting analysis showed that these antibodies did not react with classical human pancreatic lipase (HPL) or human pancreatic lipase-related protein 1 (HPLRP1) but cross-reacted with native rat PLRP2 (RPLRP2), as well as with recombinant rat and guinea-pig PLRP2 (GPLRP2). Immunoaffinity chromatography was performed on immobilized anti-recombinant HPLRP2 polyclonal antibodies to purify native HPLRP2 after conventional chromatographic steps including gel filtration and chromatrography on an anion-exchanger. The substrate specificity of HPLRP2 was investigated using various triglycerides, phospholipids and galactolipids as substrates. The lipase activity on triglycerides was inhibited by bile salts and weakly restored by colipase. The phospholipase activity of HPLRP2 on phospholipid micelles was very low. A significant level of galactolipase activity was measured using monogalactosyldiglyceride monomolecular films. These data suggest that the main physiological function of HPLRP2 is the hydrolysis of galactolipids, which are the main lipids present in vegetable food. |
Databáze: | OpenAIRE |
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