Expression of recombinant cyclooxygenase 1 in Drosophila melanogaster S2 cells transformed with human beta1,4-galactosyltransferase and Galbeta1,4-GlcNAc alpha2,6-sialyltransferase.
We examined the expression of human cyclooxygenase-1 (COX-1) in Drososphila melanogaster S2 (S2) cells transformed with cDNAs encoding beta1,4-galactosyltransferase (GalT) and Galbeta1,4-GlcNAc alpha2,6-sialyltransferase (ST). Southern blot analysis indicated that multiple copies of the glycosyltransferases genes were integrated into the S2 cell genome. A lectin blot analysis also indicated that recombinant COX-1 from S2COX-1/GalT-ST cells contained the glycan residues of beta1,4-linked galactose and alpha2,6-linked sialic acid. The specific peroxidase activity of recombinant sialylated COX-1 from S2COX-1/GalT-ST cells was 41,250 U mg(-1), indicating an increase of approximately 22% compared with a non-sialylated control (33,850 U mg(-1)) from S2COX-1 cells.
Chang
Kyung Hwa
KH
Graduate School of Biotechnology and Plant Metabolism Research Center, Kyung Hee University, Suwon, 446-701, Korea.
Lee
Jong Min
JM
Hwang-Bo
Jeon
J
Yoo
Ki Hyun
KH
Sohn
Bong Hee
BH
Yang
Jai Myung
JM
Chung
In Sik
IS
D000818:Animals; D015139:Blotting, Southern; D002460:Cell Line; D051545:Cyclooxygenase 1 / Q000302:isolation & purification / Q000378:metabolism*; D004331:Drosophila melanogaster / Q000166:cytology* / Q000201:enzymology* / Q000235:genetics; D005700:Galactosyltransferases / Q000378:metabolism*; D049750:Genome, Insect; D006801:Humans; D037102:Lectins; D009195:Peroxidase; D010957:Plasmids; D011994:Recombinant Proteins / Q000378:metabolism*; D012799:Sialyltransferases / Q000378:metabolism*; D014170:Transformation, Genetic; D000094802:beta-D-Galactoside alpha 2-6-Sialyltransferase
D037102:Lectins; D011994:Recombinant Proteins; D009195:Peroxidase; D051545:Cyclooxygenase 1; D005700:Galactosyltransferases; C493310:beta1,4-galactosyltransferase, human; D012799:Sialyltransferases; D000094802:beta-D-Galactoside alpha 2-6-Sialyltransferase