Recombinant C-terminal domain of pancreatic lipase retains full ability to bind colipase.
The organization of the pancreatic lipase in two well defined domains has been correlated to a specific function for each domain, catalytic activity for the N-terminal domain and colipase binding for the C-terminal domain. In order to see if such an organization implies that the two domains can behave as separate entities, we expressed the N- and C-terminal domains in insect cells. The recombinant proteins secreted in the cell supernatants present the expected molecular properties. However, whereas the C-terminal domain retains its function of colipase binding, the N-terminal domain appears to be unable to ensure catalysis. The lack of activity of the recombinant N-terminal domain could result either from a (partially) incorrect folding or from an incapacity to function by itself. These results suggest that, although both are structurally well defined, the two domains of the pancreatic lipase behave differently when they are expressed as separate entities.
Ayvazian
L
L
Laboratoire de Bioénergétique et Ingénierie des Protéines, UPR 9036, CNRS, Marseille, France.
Crenon
I
I
Granon
S
S
Chapus
C
C
Kerfelec
B
B
D000818:Animals; D001483:Base Sequence; D015153:Blotting, Western; D003089:Colipases / Q000378:metabolism*; D018076:DNA, Complementary / Q000737:chemistry; D004591:Electrophoresis, Polyacrylamide Gel; D008049:Lipase / Q000378:metabolism*; D010179:Pancreas / Q000201:enzymology*; D011994:Recombinant Proteins / Q000737:chemistry / Q000378:metabolism; D013552:Swine
D003089:Colipases; D018076:DNA, Complementary; D011994:Recombinant Proteins; D008049:Lipase