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These lipid modifications are widely accepted to be important for the biological activity and secretion of Wnt proteins.
Binding of different members of the Wnt-family to certain members of the Fzd family can initiate signaling by one of several distinct pathways.
In the "canonical pathway," activation of the signaling molecule, Disheveled, leads to the inactivation of glycogen synthase kinase-3 (GSK3P), a cytoplasmic serine-threonine kinase.
Optionally, the peptide may then be contacted with a glycosyltransferase specific for a galactose acceptor moiety and an sialic acid acceptor moiety, in the presence of sialic acid-nucleotide donor molecules to ensure complete the SA capping of all of the glycan molecules.
The trimannose core is present in the vast majority of the N-linked glycans made by insect cells, and sometimes an antennary Glc NAc and/or fucose residue(s) are also present.
The conjugated moiety may be a PEG molecule, another peptide, a small molecule such as a drug, among others.
This embodiment therefore is useful for conjugating the peptide to PEG molecules that will slow the clearance of the peptide from the patient's bloodstream, to peptides that will target both peptides to a specific tissue or cell, or to another peptide of complementary therapeutic use.
This example is particularly advantageous for human peptides expressed in heterologous cellular expression systems.
By remodeling the N-linked glycan structures to this configuration, the peptide can be made less immunogenic in a human patient, and/or more stable, among others.
This example is particularly advantageous for recombinant antibody molecules expressed in heterologous cellular systems.
When the antibody molecule includes a Fc-mediated cellular cytotoxicity, it is known that the presence of bisected oligosaccharides linked the Fc domain dramatically increased antibody-dependent cellular cytotoxicity.
In one exemplary embodiment, the O-linked glycan on a peptide expressed in an insect cell is remodeled to a humanized glycan by contacting the glycans with a glycosyltransferase specific for a Gal NAc acceptor molecule and a galactose donor molecule in the presence of nucleotide-Gal; and then contacting the glycans with a glycosyltransferase specific for a Gal acceptor molecule and a SA donor molecule in the presence of nucleotide-SA.Tags: Adult Dating, affair dating, sex dating