Cloning and expression in insect cells of two pancreatic lipases and a procolipase from Myocastor coypus.
The physiological role of pancreatic lipases has traditionally been assigned solely to triacylglyceride metabolism, while the digestion of phospholipids requires the presence of the pancreatic phospholipase A2, a 14-kDa enzyme unrelated to pancreatic lipases. However, in the guinea pig, it was observed that the pancreatic phospholipase A2 was absent and that a guinea pig pancreatic-lipase-related protein 2 (GPL-RP2) was responsible for phospholipase activity, in contrast to the situation observed in other mammalian species. As the guinea pig is a member of the hystricomorph rodents, it was of interest to investigate if other species within this evolutionary suborder display similar characteristics. The coypu (Myocastor coypus) also a member of the hystricomorph rodents, was chosen for further investigations. The cDNAs encoding two pancreatic lipases and a procolipase from the coypu were cloned, expressed and characterized. One lipase, CoPL-RP2, was identified as belonging to the RP2 subfamily, while the second, CoPL, was found to belong to the classical pancreatic lipase subfamily. Enzymic characterization and sequence data suggest a role for coypu colipase as a specific cofactor for CoPL, while this coypu colipase cannot be an important cofactor for CoPL-RP2 in vivo. Also, the new lipase cDNA sequences were used in a phylogentic analysis to reinvestigate the taxonomical position of the hystricomorph rodents (e.g. coypu and guinea pig) with respect to the myomorph rodents (e.g. rat and mouse).
Thirstrup
K
K
Carrière
F
F
Hjorth
S A
SA
Rasmussen
P B
PB
Nielsen
P F
PF
Ladefoged
C
C
Thim
L
L
Boel
E
E
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D003001:Cloning, Molecular; D003089:Colipases / Q000235:genetics*; D004247:DNA; D004792:Enzyme Precursors; D008049:Lipase / Q000235:genetics*; D008969:Molecular Sequence Data; D017924:Nucleopolyhedroviruses / Q000235:genetics; D010179:Pancreas / Q000201:enzymology*; D010802:Phylogeny; D011498:Protein Precursors / Q000235:genetics*; D012377:Rodentia; D018411:Spodoptera
D003089:Colipases; D004792:Enzyme Precursors; D011498:Protein Precursors; C037107:procolipase; D004247:DNA; D008049:Lipase