Site-Specific Transglutaminase-Mediated Conjugation of Interferon α-2b at Glutamine or Lysine Residues
| Autor: | Isabella Monia Montagner, Antonio Rosato, Angelo Fontana, Antonella Grigoletto, Barbara Spolaore, Gianfranco Pasut, Samanta Raboni, Abhijeet Satwekar, Anna Mero |
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| Jazyk: | angličtina |
| Rok vydání: | 2016 |
| Předmět: |
Models
Molecular 0301 basic medicine Tissue transglutaminase Glutamine Lysine Biomedical Engineering Pharmaceutical Science Alpha interferon Bioengineering Interferon alpha-2 Antiviral Agents Protein Structure Secondary Polyethylene Glycols Substrate Specificity law.invention Interferon α-2b 03 medical and health sciences transglutaminase law protein conjugation Animals Humans Amino Acid Sequence Binding site Peptide sequence Interferon α-2b protein conjugation transglutaminase PEGylation limited proteolysis Pharmacology Binding Sites Transglutaminases integumentary system biology Chemistry Organic Chemistry PEGylation Interferon-alpha Vesiculovirus Ligand (biochemistry) Molecular biology Recombinant Proteins Rats Molecular Weight 030104 developmental biology Biochemistry biology.protein Recombinant DNA Biotechnology limited proteolysis |
| Popis: | Interferon α (IFN α) subtypes are important protein drugs that have been used to treat infectious diseases and cancers. Here, we studied the reactivity of IFN α-2b to microbial transglutaminase (TGase) with the aim of obtaining a site-specific conjugation of this protein drug. Interestingly, TGase allowed the production of two monoderivatized isomers of IFN with high yields. Characterization by mass spectrometry of the two conjugates indicated that they are exclusively modified at the level of Gln101 if the protein is reacted in the presence of an amino-containing ligand (i.e., dansylcadaverine) or at the level of Lys164 if a glutamine-containing molecule is used (i.e., carbobenzoxy-l-glutaminyl-glycine, ZQG). We explained the extraordinary specificity of the TGase-mediated reaction on the basis of the conformational features of IFN. Indeed, among the 10 Lys and 12 Gln residues of the protein, only Gln101 and Lys164 are located in highly flexible protein regions. The TGase-mediated derivatization of IFN was then applied to the production of IFN derivatives conjugated to a 20 kDa polyethylene glycol (PEG), using PEG-NH |
| Databáze: | OpenAIRE |
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