Molecular cloning and expression of rabbit pancreatic cholesterol esterase.
Rabbit pancreatic cholesterol esterase (CEase, carboxyl ester lipase, EC 3.1.1.3) has been cloned from a lambda gt11 library of adult rabbit pancreatic cDNA. The open reading frame consists of 1788 nucleotides which encodes 576 amino acids of the functional protein and a 20 amino acid leader peptide. When compared to other species, the greatest homology is observed between residues 82-248 with little or no homology at the C-terminal end where proline-glutamate-serine-threonine (PEST) segments are a characteristic feature of the human CEase. Rabbit CEase (RCEase) retains the active-site serine (gxsxg), the active-site histidine and the tentative heparin binding site (KKRCLQ) at similar positions in comparison to pancreatic CEases of other species. When rabbit CEase cDNA is expressed in monkey kidney (COS-7) cells, enzymatic hydrolytic activity is detected in the growth medium as is a 67 kDa protein by Western blotting with polyclonal anti-CEase antibody. Northern blot analysis shows two mRNA (2.2 and 3.2 kb) species.
Colwell
N S
NS
Geriatric Research Education and Clinical Center, V.A. Medical Center, St. Louis, MO.
Aleman-Gomez
J A
JA
Kumar
B V
BV
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D002417:Cattle; D003001:Cloning, Molecular; D004247:DNA / Q000235:genetics; D015723:Gene Library; D006801:Humans; D008969:Molecular Sequence Data; D016366:Open Reading Frames; D010179:Pancreas / Q000201:enzymology*; D021382:Protein Sorting Signals / Q000235:genetics; D011817:Rabbits; D051381:Rats; D017386:Sequence Homology, Amino Acid; D002787:Sterol Esterase / Q000235:genetics*
D021382:Protein Sorting Signals; D004247:DNA; D002787:Sterol Esterase