Engineering potent long-acting variants of the Wnt inhibitor DKK2
| Autor: | Joseph W Arndt, Renée I Shapiro, Abhishek Kulkarni, Blake Pepinsky, Joseph Amatucci, Nels E Pederson, Andreas Lehmann, Dingyi Wen, Joshua W Mugford, Joseph Worrall, Brenda K. Minesinger, Richelle Sopko, Mia Rushe |
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| Rok vydání: | 2017 |
| Předmět: |
0301 basic medicine
Recombinant Fusion Proteins Bioengineering Context (language use) Protein Engineering Biochemistry Dickkopf-2 Mice 03 medical and health sciences chemistry.chemical_compound Animals Humans Wnt Signaling Pathway Molecular Biology Serum Albumin Wnt signaling pathway Wild type LRP6 Heparan sulfate Protein engineering Wnt signaling tissue injury Cell biology Wnt Proteins 030104 developmental biology chemistry human serum albumin Low Density Lipoprotein Receptor-Related Protein-6 Intercellular Signaling Peptides and Proteins Phosphorylation Original Article Heparan sulfate binding heparan sulfate Biotechnology |
| Zdroj: | Protein Engineering, Design and Selection |
| ISSN: | 1741-0134 1741-0126 |
| DOI: | 10.1093/protein/gzx007 |
| Popis: | Wnt signaling pathways are required for a wide variety of biological processes ranging from embryonic development to tissue repair and regeneration. Dickkopf-2 (DKK2) is classically defined as a canonical Wnt inhibitor, though it may play a role in activating non-canonical Wnt pathways in the context of endothelial network formation after acute injury. Here we report the discovery of a fusion partner for a DKK2 polypeptide that significantly improves the expression, biochemical properties and pharmacokinetics (PK) of the DKK2 polypeptide. Specifically, human serum albumin (HSA) was identified as a highly effective fusion partner. Substitution of selected amino acid residues in DKK2 designed to decrease heparan sulfate binding by HSA-DKK2 variants, further improved the PK properties of the molecule in rodents. The HSA-DKK2 variants were monomeric, as thermally stable as wild type, and active as measured by their ability to bind to and prevent phosphorylation of the Wnt coreceptor LRP6. Our engineering efforts resulted in potent long-lived variants of the canonical Wnt inhibitor DKK2, applicable for Wnt pathway manipulation either by systematic delivery or focused administration at sites of tissue injury. |
| Databáze: | OpenAIRE |
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