Addition of G418 and other aminoglycoside antibiotics to mammalian cells results in the release of GPI-anchored proteins.
Resistance to the neomycin analogue G418 forms the basis of a dominant marker selection system for mammalian cells transfected with the bacterial neomycin gene. We found that COS-1 cells stably transfected with the neomycin resistance gene had a greater than 50% reduction in cell-associated glycosylphosphatidylinositol (GPI)-anchored alkaline phosphatase (AP). A similarly reduced amount of AP was also observed in wild-type COS-1 cells incubated in the presence of G418 or other aminoglycoside antibiotics. The AP was released from cells into the culture supernatant in its GPI-anchored form. Our data suggest that the G418-induced reduction of AP involves a vesiculation process of COS-1 cells.
Küng
M
M
Institute of Biochemistry and Molecular Biology, University of Bern, Switzerland.
Stadelmann
B
B
Brodbeck
U
U
Bütikofer
P
P
D000469:Alkaline Phosphatase / Q000187:drug effects / Q000235:genetics / Q000378:metabolism; D000818:Animals; D000900:Anti-Bacterial Agents / Q000494:pharmacology*; D019556:COS Cells / Q000187:drug effects / Q000201:enzymology; D002522:Chlorocebus aethiops; D004305:Dose-Response Relationship, Drug; D004789:Enzyme Activation; D004912:Erythrocytes / Q000187:drug effects; D005839:Gentamicins / Q000494:pharmacology*; D017261:Glycosylphosphatidylinositols / Q000378:metabolism*; D006801:Humans; D008565:Membrane Proteins / Q000187:drug effects / Q000378:metabolism*; D013997:Time Factors; D014162:Transfection
D000900:Anti-Bacterial Agents; D005839:Gentamicins; D017261:Glycosylphosphatidylinositols; D008565:Membrane Proteins; C010680:antibiotic G 418; D000469:Alkaline Phosphatase