Human pancreatic procolipase expressed in insect cells: purification and characterization.
A cDNA clone encoding human pancreatic procolipase was incorporated into a recombinant baculovirus. Spodoptera frugiperda insect cells infected with the recombinant baculovirus secreted procolipase into the medium, which could be isolated in a single step by immunoaffinity chromatography. The highly purified protein reactivated human pancreatic lipase in a concentration-dependent fashion and was efficiently converted to colipase by limited trypsin digestion. This expression system is suitable for producing amounts of procolipase adequate for biophysical studies.
Lowe
M E
ME
Department of Pediatrics, Washington University School of Medicine, St. Louis, Missouri 63110.
D000818:Animals; D016367:Baculoviridae / Q000235:genetics; D002478:Cells, Cultured; D002850:Chromatography, Gel; D003089:Colipases / Q000235:genetics / Q000302:isolation & purification* / Q000378:metabolism*; D004792:Enzyme Precursors; D015870:Gene Expression; D006801:Humans; D008049:Lipase / Q000378:metabolism*; D010179:Pancreas / Q000201:enzymology; D011498:Protein Precursors / Q000235:genetics / Q000302:isolation & purification* / Q000378:metabolism*; D011499:Protein Processing, Post-Translational; D011995:Recombination, Genetic; D018411:Spodoptera / Q000378:metabolism; D014357:Trypsin
D003089:Colipases; D004792:Enzyme Precursors; D011498:Protein Precursors; C037107:procolipase; D008049:Lipase; D014357:Trypsin