Insect cell hosts for baculovirus expression vectors contain endogenous exoglycosidase activity.
Four different insect cell lines that can be used as hosts for baculovirus infection were assayed for the presence of endogenous exoglycosidases. All four cell lines, derived from Spodoptera frugiperda, Trichoplusia ni, Bombyx mori, or Malacosoma disstria, contained N-acetyl-beta-glucosaminidase, N-acetyl-beta-galactosaminidase, beta-galactosidase, and sialidase activities. Exoglycosidase activities were found in cell lysates as well as cell-free supernatants from uninfected and wild-type baculovirus infected cells. Oligosaccharide analysis of cellular glycoproteins using lectins recognizing Gal beta 1, 3GalNAc, Gal beta 1, 4GlcNAc, and NeuAc alpha 2,6Gal demonstrated that only Gal beta 1,3GalNAc was present. The demonstration that these cells contain exoglycosidases raises the possibility that the oligosaccharides of baculovirus-expressed glycoproteins are subject to enzymatic degradation.
Licari
P J
PJ
Department of Chemical Engineering, California Institute of Technology, Pasadena 91125.
Jarvis
D L
DL
Bailey
J E
JE
D000118:Acetylglucosaminidase / Q000378:metabolism; D000818:Animals; D016367:Baculoviridae / Q000235:genetics*; D002240:Carbohydrate Sequence; D002478:Cells, Cultured; D006026:Glycoside Hydrolases / Q000378:metabolism*; D006596:Hexosaminidases / Q000378:metabolism; D037102:Lectins / Q000378:metabolism; D008969:Molecular Sequence Data; D009036:Moths / Q000166:cytology / Q000201:enzymology* / Q000821:virology; D009439:Neuraminidase / Q000378:metabolism; D019887:Peanut Agglutinin; D001616:beta-Galactosidase / Q000378:metabolism; D048828:beta-N-Acetyl-Galactosaminidase; D001619:beta-N-Acetylhexosaminidases / Q000378:metabolism
D037102:Lectins; D019887:Peanut Agglutinin; D006026:Glycoside Hydrolases; D006596:Hexosaminidases; D009439:Neuraminidase; D001616:beta-Galactosidase; D000118:Acetylglucosaminidase; D001619:beta-N-Acetylhexosaminidases; D048828:beta-N-Acetyl-Galactosaminidase