Document Detail


Stable expression of enzymatically active human pancreatic lipase in V79 cells: purification and characterization of the recombinant enzyme.
MedLine Citation:
PMID:  8013095     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
The serum activity of human pancreatic lipase (HPL; EC 3.1.1.3), the main lipolytic enzyme secreted by the pancreas, is a valuable marker of pancreatic disorders. However, determining lipase activity in human serum is difficult because the substrates used vary in their lipase specificity. A lipase reference material is therefore needed. We describe the production of recombinant HPL (rHPL) in V79 Chinese hamster lung cells. A cDNA encoding the sequence of HPL was subcloned into the pcDNAI eukaryotic expression vector, which was then used to transfect V79 cells. The 50-kDa purified recombinant enzyme is fully active, is glycosylated, and has a pI of 7.0. The catalytic properties of rHPL, determined by using triolein as the substrate, were similar to those of the native enzyme. The V79rHPL cell line we developed might be useful for the production of rHPL suitable for the preparation of a reference material.
Authors:
F Canalias; A Visvikis; C Thioudellet; G Siest
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Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't    
Journal Detail:
Title:  Clinical chemistry     Volume:  40     ISSN:  0009-9147     ISO Abbreviation:  Clin. Chem.     Publication Date:  1994 Jul 
Date Detail:
Created Date:  1994-07-28     Completed Date:  1994-07-28     Revised Date:  2006-11-15    
Medline Journal Info:
Nlm Unique ID:  9421549     Medline TA:  Clin Chem     Country:  UNITED STATES    
Other Details:
Languages:  eng     Pagination:  1251-7     Citation Subset:  IM    
Affiliation:
Centre du Médicament, URA CNRS 597 30, Nancy, France.
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MeSH Terms
Descriptor/Qualifier:
Animals
Base Sequence
Cell Line
Cloning, Molecular
Cricetinae
Cricetulus
Culture Media
DNA, Complementary / genetics
Gene Expression*
Glycosylation
Isoelectric Point
Kinetics
Lipase / genetics*,  isolation & purification,  metabolism
Lung
Molecular Sequence Data
Pancreas / enzymology*
Recombinant Proteins / isolation & purification,  metabolism
Transfection
Chemical
Reg. No./Substance:
0/Culture Media; 0/DNA, Complementary; 0/Recombinant Proteins; EC 3.1.1.3/Lipase

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