Immunochemical studies of pancreatic colipase-lipase interaction employing immobilized synthetic peptides.
In view to study the possible participation of the sequence portions of colipase including or close to the free carboxyl groups at positions 15 and/or 72 to the binding with pancreatic lipase, we have used three synthetic peptides matching portions 8-16, 59-67 and 67-72 of the amino acid sequence. Polyclonal rabbit anticolipase immune serum, which cross-reacts with peptides in ELISA, was fractionated on columns of peptide coupled to Sepharose. Of the three fractions of antibodies, only that interacting with peptide 8-16 had the capacity to inhibit colipase-dependent lipase activity by specifically preventing the association of lipase with its protein cofactor previously bound to lipid. We conclude that the region spanning residues 8-16 of colipase is of importance for colipase-lipase interaction in the active complex formed at interface.
Rugani
N
N
Institut de Chimie Biologique, Faculté Saint-Charles, Marseille, France.
de la Fournière
L
L
Julien
R
R
Sarda
L
L
Rathelot
J
J
D000595:Amino Acid Sequence; D000818:Animals; D000906:Antibodies; D002846:Chromatography, Affinity; D003089:Colipases / Q000302:isolation & purification / Q000378:metabolism*; D003429:Cross Reactions; D004797:Enzyme-Linked Immunosorbent Assay; D006736:Horses; D007700:Kinetics; D008049:Lipase / Q000302:isolation & purification / Q000378:metabolism*; D008969:Molecular Sequence Data; D009842:Oligopeptides / Q000138:chemical synthesis / Q000276:immunology; D010179:Pancreas / Q000201:enzymology*; D013552:Swine
D000906:Antibodies; D003089:Colipases; D009842:Oligopeptides; D008049:Lipase