We designed three types of RGD-containing barnacle adhesive proteins using self-assembling peptides. In the present study, we designed a peptide of which the self-assembling peptide Arg-Arg-Lys-Ser-Tyr-Ser-Gly-Ile-Leu-Gly-Asp-Leu-Ile-Gln-Ala-Val-Ile-Arg-Tyr-Tyr (RY) was modified with the cell adhesion peptide RGDS. Three types of molecules were designed: R-Y-RGDS with RGDS modified at the C-terminus of R-Y, RGDS-R-Y with RGDS modified at the N-terminus of R-Y, and R-RGD-Y with RGD introduced into the sequence of R-Y. R-RGD-Y was replaced leucine with arginine in the center of R-Y; -LGD- sequence to -RGD-. Three types of RGD-containing peptides were synthesized by solidphase peptide synthesis, and the secondary structures of these peptides were analyzed by CD and FT-IR spectroscopy. The mechanical properties of peptide hydrogels were characterized by a rheometer. We discuss the correlation between the peptide conformation, and cell attachment and cell spreading activity from the viewpoint of developing effective tissue engineering scaffolds. We created a peptide-coated cell culture substrate by coating peptides on a polystyrene plate. They significantly facilitated cell adhesion and spreading compared to a non-coated substrate. When the RGDS sequence was modified at N- or C-terminal of R-Y, it was found that the self-assembling ability was dependent on the strongly affects hydrogel formation and cell adhesion caused by its secondary structure.

Design of RGDS-immobilized self-assembling peptide from barnacle protein

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