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Modification of a recombinant GPI‐anchored metalloproteinase for secretion alters the protein glycosylation
Author(s) -
Morrison Charlotte J.,
Easton Richard L.,
Morris Howard R.,
McMaster W. Robert,
Piret James M.,
Dell Anne
Publication year - 2000
Publication title -
biotechnology and bioengineering
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.136
H-Index - 189
eISSN - 1097-0290
pISSN - 0006-3592
DOI - 10.1002/(sici)1097-0290(20000520)68:4<407::aid-bit6>3.0.co;2-s
Subject(s) - glycosylation , secretion , recombinant dna , metalloproteinase , chemistry , biochemistry , secretory protein , posttranslational modification , microbiology and biotechnology , biology , matrix metalloproteinase , enzyme , gene
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. © 2000 John Wiley & Sons, Inc. Biotechnol Bioeng 68: 407–421, 2000.