Modified expression of coagulation factor VIII by addition of a glycosylation site at the N terminus of the protein.
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Srour MA, Grupp J, Aburubaiha Z, Albert T, Brondke H, Oldenburg J, Schwaab R
Modified expression of coagulation factor VIII by addition of a glycosylation site at the N terminus of the protein.
Ann Hematol. 2008 Feb;87(2):107-12. Epub 2007 Sep 26.
- PubMed ID
- 17899080 [ View in PubMed]
- Abstract
Recently, it was shown that glycoproteins with N-glycans close to the NH(2) terminus can directly enter the calnexin/calreticulin cycle and bypass BiP binding. This should allow efficient secretion of glycoproteins such as factor VIII (FVIII) whose secretion is negatively affected by BiP interaction. Examination of the glycosylation pattern of the NH(2) terminus of FV and FVIII revealed N-glycans at positions 23 and 27 in FV and at position 41 in FVIII. To improve FVIII secretion, a 14-amino-acid-long polypeptide with (G3) or without (G0; control) three N-linked glycosylation consensus sites was inserted upstream of the NH(2) terminus of a B-domain deleted FVIII protein. Expression of G3- and G0-constructs in three different cell lines resulted in the same or even higher expression rate of protein as found for the B-domain deleted FVIII. However, as demonstrated by Western blot analysis, the G3- as well as the G0-protein variants were mainly retained inside the cells in similar amounts. Thus, glycosylation alone does not automatically lead to higher secretion rates, but must be in context to the normal structure of the FVIII protein.
DrugBank Data that Cites this Article
- Drug Targets
Drug Target Kind Organism Pharmacological Action Actions Antihemophilic factor, human recombinant Calnexin Protein Humans UnknownChaperoneDetails Antihemophilic factor, human recombinant Calreticulin Protein Humans UnknownChaperoneDetails Lonoctocog alfa Calnexin Protein Humans UnknownChaperoneDetails Lonoctocog alfa Calreticulin Protein Humans UnknownChaperoneDetails Moroctocog alfa Calnexin Protein Humans UnknownChaperoneDetails Moroctocog alfa Calreticulin Protein Humans UnknownChaperoneDetails