Expression of egasyn-esterase in mammalian cells. Sequestration in the endoplasmic reticulum and complexation with beta-glucuronidase.
Mouse egasyn cDNA was inserted into expression vector pCDpoly and transfected into mammalian cell lines. Transfected human HepG2 cells, monkey COS-1 cells, and mouse L cells expressed egasyn-esterase catalytic activity. Within COS-1 cells, egasyn was localized to the endoplasmic reticulum. Although individual cells produced large amounts of egasyn, no secretion was observed. No beta-glucuronidase-egasyn complexes were formed in transfected HepG2 or COS-1 cells. However, these complexes were readily detected in transfected L cells. Although the signal for retention of egasyn in the endoplasmic reticulum appears to be species independent, the signal for association with beta-glucuronidase is species restricted.
Novak
E K
EK
Department of Molecular and Cellular Biology, Roswell Park Cancer Institute, Buffalo, New York 14263.
Baumann
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H
Ovnic
M
M
Swank
R T
RT
D000818:Animals; D015152:Blotting, Northern; D002265:Carboxylic Ester Hydrolases / Q000235:genetics*; D004247:DNA / Q000235:genetics* / Q000378:metabolism; D004591:Electrophoresis, Polyacrylamide Gel; D004950:Esterases / Q000235:genetics* / Q000378:metabolism; D005455:Fluorescent Antibody Technique; D005786:Gene Expression Regulation; D005966:Glucuronidase / Q000235:genetics / Q000378:metabolism; D008562:Membrane Glycoproteins / Q000235:genetics*; D051379:Mice; D010957:Plasmids; D012333:RNA, Messenger / Q000032:analysis; D014162:Transfection
D008562:Membrane Glycoproteins; D012333:RNA, Messenger; D004247:DNA; D004950:Esterases; D002265:Carboxylic Ester Hydrolases; C009751:egasyn; D005966:Glucuronidase