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.
Department of Pediatrics, Washington University School of Medicine, St. Louis, Missouri 63110.
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Lowe
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