cDNA cloning of human-milk bile-salt-stimulated lipase and evidence for its identity to pancreatic carboxylic ester hydrolase.
We have isolated and sequenced cDNA clones covering the entire coding sequence of human-milk bile-salt-stimulated lipase, as well as 996 nucleotides of the 3' end of the pancreatic enzyme carboxylic ester hydrolase. The deduced amino acid sequence of the lipase starts with a 23-residue leader peptide. The open reading frame continues with 722 amino acid residues. The sequence contains in the C-terminal part a proline-rich repeat, 16 repeats of 11 amino acid residues each. The mRNA was estimated to be approximately 2500 nucleotides from Northern blot and of similar size in mammary and pancreatic tissues. Data obtained indicate that the lipase and the carboxylesterase are identical and coded for by the same gene. The cDNA is 2428 bases long, which indicates that a near full-length copy of the transcript has been isolated. Comparisons with other enzymes show that the lipase is a new member of the supergene family of serine hydrolases. It is not only closely related (and in its N-terminal half virtually identical) to lysophospholipase from rat pancreas and cholesterol esterase from bovine pancreas, but also shows a high degree of similarity to several esterases, e.g. acetylcholine esterase. In contrast, no such similarity could be found to typical lipases.
Department of Molecular Biology, University of Göteborg, Sweden.
J
J
Nilsson
L
L
Bläckberg
P
P
Carlsson
S
S
Enerbäck
O
O
Hernell
G
G
Bjursell
D000273:Adipose Tissue / Q000201:enzymology; D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D001647:Bile Acids and Salts / Q000494:pharmacology; D001940:Breast / Q000201:enzymology; D043182:Carboxylesterase; D002265:Carboxylic Ester Hydrolases / Q000235:genetics*; D003001:Cloning, Molecular; D004247:DNA / Q000235:genetics* / Q000302:isolation & purification; D005260:Female; D006801:Humans; D008049:Lipase / Q000235:genetics* / Q000378:metabolism; D008895:Milk, Human / Q000201:enzymology*; D008969:Molecular Sequence Data; D010179:Pancreas / Q000201:enzymology*; D010446:Peptide Fragments / Q000302:isolation & purification; D011247:Pregnancy; D021382:Protein Sorting Signals / Q000235:genetics; D012313:RNA / Q000235:genetics / Q000302:isolation & purification; D012333:RNA, Messenger / Q000235:genetics; D012689:Sequence Homology, Nucleic Acid
D001647:Bile Acids and Salts; D010446:Peptide Fragments; D021382:Protein Sorting Signals; D012333:RNA, Messenger; D012313:RNA; D004247:DNA; D002265:Carboxylic Ester Hydrolases; D043182:Carboxylesterase; D008049:Lipase