Expression of the murine prostaglandin (PGH) synthase-1 and PGH synthase-2 isozymes in cos-1 cells.
Plasmid vectors were constructed which allowed expression of the mouse prostaglandin endoperoxide (PGH) synthase-1 and PGH synthase-2 isozymes in cos-1 cells. Efficient expression of the PGHS-2 isozyme required the truncation of the entire 3'-untranslated region of the PGHS-2 cDNA, possibly due to the presence of multiple AUUUA sequences which may destabilize the PGHS-2 mRNA. The length of the 3'-untranslated regions of the murine and ovine PGHS-1 isozymes, which do not contain AUUUA sequences, did not affect the efficiency of expression of these proteins. The murine PGHS-2 isozyme catalyzes the same cyclooxygenase and hydroperoxidase activities as the ovine and murine PGHS-1 isozymes. The maximal activities of the mouse enzymes expressed in cos-1 cells was about equal, but both were only about a third that seen with the sheep enzyme. Whether this reflects differences in the turnover rate of the mouse and sheep enzymes, or differences in the efficiency of expression in cos-1 cells is not known.
Meade
E A
EA
Department of Biochemistry, Michigan State University, East Lansing 48824.
Smith
W L
WL
DeWitt
D L
DL
D000818:Animals; D002460:Cell Line; D003001:Cloning, Molecular; D004247:DNA / Q000235:genetics; D015870:Gene Expression; D005822:Genetic Vectors; D007527:Isoenzymes / Q000235:genetics* / Q000378:metabolism; D051379:Mice; D011451:Prostaglandin-Endoperoxide Synthases / Q000235:genetics* / Q000378:metabolism; D015183:Restriction Mapping; D012756:Sheep
D007527:Isoenzymes; D004247:DNA; D011451:Prostaglandin-Endoperoxide Synthases