title: Directional BMP-2 for functionalization of titanium surfaces.
abstract: Efficient immobilization of biomacromolecules on material surfaces is a key to development in areas of regenerative medicine and tissue engineering. However, strong and irreversible immobilization of cytokines on surfaces often diminishes their biological functionality. A destructive hydrophobic interaction between the material surface and the biomolecule may underlie this inactivation. Alternatively, dissociation of the cytokine from the material may be necessary for signal transduction. Here we propose a new method for immobilizing cytokines on material surfaces: a material-binding artificial peptide is used to mediate reversible interaction between the cytokine and the material surface. We created artificial proteins that contained three copies of a Ti-binding motif, and fused them to the N-terminal of BMP-2. The engineered BMP-2 showed reversible binding to Ti surfaces and induced BMP signaling activity. When a hydrophobic protein devoid of the Ti-binding motif was fused to BMP-2, the protein tightly bound to Ti surfaces but showed little BMP activity, confirming the importance of the mode of immobilization.
authors.affiliation: Department of Protein Engineering, Cancer Institute, Japanese Foundation for Cancer Research, 3-10-6 Ariake, Koto-ku, Tokyo 135-8550, Japan.
authors.forename: Kenji
authors.initials: K
authors.lastname: Kashiwagi
authors.forename: Toru
authors.initials: T
authors.lastname: Tsuji
authors.forename: Kiyotaka
authors.initials: K
authors.lastname: Shiba
mesh_terms: D000327:Adsorption / Q000187:drug effects; D000469:Alkaline Phosphatase / Q000378:metabolism; D000595:Amino Acid Sequence; D000818:Animals; D055396:Bone Morphogenetic Protein 2 / Q000737:chemistry / Q000494:pharmacology*; D052005:Bone Morphogenetic Protein Receptors, Type I / Q000378:metabolism; D002454:Cell Differentiation / Q000187:drug effects; D002460:Cell Line; D006801:Humans; D007150:Immunohistochemistry; D051379:Mice; D008969:Molecular Sequence Data; D018485:Muscle Fibers, Skeletal / Q000166:cytology / Q000187:drug effects / Q000201:enzymology; D010455:Peptides / Q000737:chemistry / Q000494:pharmacology; D011993:Recombinant Fusion Proteins / Q000737:chemistry; D013499:Surface Properties / Q000187:drug effects; D014025:Titanium / Q000737:chemistry*
chemical_list: D055396:Bone Morphogenetic Protein 2; D010455:Peptides; D011993:Recombinant Fusion Proteins; D014025:Titanium; D052005:Bone Morphogenetic Protein Receptors, Type I; D000469:Alkaline Phosphatase