title: Modification of a recombinant GPI-anchored metalloproteinase for secretion alters the protein glycosylation.
abstract: The N-linked glycans of recombinant leishmanolysin (GP63) expressed as a glycosylphosphatidylinositol (GPI)-anchored membrane protein or modified for secretion in Chinese hamster ovary (CHO) cells were analyzed by fast atom bombardment-mass spectrometry (FAB-MS). The glycans isolated from both membrane and secreted protein were predominantly complex biantennary structures. However other aspects of the glycan profiles showed striking differences. The degree of sialylation of the membrane form was greatly reduced and the core fucosylation of biantennary structures was increased compared to the secreted form. Glycans isolated from membrane expressed protein also contained a higher proportion of lactosamine repeats. Residence times in the secretory pathway were similar for both secreted and membrane protein. Glycosylation differences may therefore be due to differences in protein conformation and accessibility to glycosyltransferases or glycosidases. These differences in glycosylation represent an important factor when considering modifying membrane expressed proteins for secreted production.
authors.affiliation: Biotechnology Laboratory, University of British Columbia, Vancouver, British Columbia, Canada.
authors.forename: C J
authors.initials: CJ
authors.lastname: Morrison
authors.forename: R L
authors.initials: RL
authors.lastname: Easton
authors.forename: H R
authors.initials: HR
authors.lastname: Morris
authors.forename: W R
authors.initials: WR
authors.lastname: McMaster
authors.forename: J M
authors.initials: JM
authors.lastname: Piret
authors.forename: A
authors.initials: A
authors.lastname: Dell
mesh_terms: D000818:Animals; D001709:Biotechnology / Q000379:methods*; D016466:CHO Cells; D006224:Cricetinae; D006031:Glycosylation; D017261:Glycosylphosphatidylinositols; D008666:Metalloendopeptidases / Q000235:genetics* / Q000378:metabolism*; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism
chemical_list: D017261:Glycosylphosphatidylinositols; D011994:Recombinant Proteins; D008666:Metalloendopeptidases