The function of the human interferon-beta 1a glycan determined in vivo.
Recombinant human interferon-beta (rhIFN-beta) is the leading therapeutic intervention shown to change the cause of relapsing-remitting multiple sclerosis, and both a nonglycosylated and a significantly more active glycosylated variant of rhIFN-beta are used in treatment. This study investigates the function of the rhIFN-beta1a glycan moiety and its individual carbohydrate residues, using the myxovirus resistance (Mx) mRNA as a biomarker in Mx-congenic mice. We showed that the Mx mRNA level in blood leukocytes peaked 3 h after s.c. administration of rhIFN-beta1a. In addition, a clear dose-response relationship was confirmed, and the Mx response was shown to be receptor-mediated. Using specific glycosidases, different glycosylation analogs of rhIFN-beta1a were obtained, and their activities were determined. The glycosylated rhIFN-beta1a showed significantly higher activity than its deglycosylated counterpart, due to a protein stabilization/solubilization effect of the glycan. It is interesting to note that the terminating sialic acids were essential for these effects. Conclusively, the structure/bioactivity relationship of rhIFN-beta1a was determined in vivo, and it provided a novel insight into the role of the rhIFN-beta1a glycan and its carbohydrate residues. The possibilities of improving the pharmacological properties of rhIFN-beta1a using glycoengineering are discussed.
Dissing-Olesen
Lasse
L
Medical Biotechnology Center, University of Southern Denmark, Winsløwparken 25, 2, DK-5000 Odense C, Denmark. ldolesen@health.sdu.dk
Thaysen-Andersen
Morten
M
Meldgaard
Michael
M
Højrup
Peter
P
Finsen
Bente
B
D000818:Animals; D004355:Drug Stability; D019204:GTP-Binding Proteins / Q000737:chemistry / Q000235:genetics; D006023:Glycoproteins / Q000737:chemistry / Q000378:metabolism / Q000502:physiology*; D006031:Glycosylation; D006801:Humans; D007370:Interferon Type I / Q000494:pharmacology; D000068556:Interferon beta-1a; D016899:Interferon-beta / Q000737:chemistry / Q000378:metabolism / Q000502:physiology*; D051379:Mice; D020297:Mice, Congenic; D008810:Mice, Inbred C57BL; D018345:Mice, Knockout; D064547:Myxovirus Resistance Proteins; D011134:Polysaccharides / Q000737:chemistry / Q000378:metabolism / Q000502:physiology*; D011994:Recombinant Proteins; D012794:Sialic Acids / Q000502:physiology; D013329:Structure-Activity Relationship
D006023:Glycoproteins; D007370:Interferon Type I; C579070:Mx1 protein, mouse; D064547:Myxovirus Resistance Proteins; D011134:Polysaccharides; D011994:Recombinant Proteins; D012794:Sialic Acids; D016899:Interferon-beta; D019204:GTP-Binding Proteins; D000068556:Interferon beta-1a