Solid State Self-Assembly Mechanism of RADA16-I Designer Peptide

We report that synthetic RADA16-I peptide transforms to beta-strand secondary structure and develops intermolecular organization into beta-sheets when stored in the solid state at room temperature. Secondary structural changes were probed using solid state nuclear magnetic resonance spectroscopy (ssNMR) and Fourier transform infrared spectroscopy (FTIR). Intermolecular organization was analyzed via wide-angle X-ray diffraction (WAXD). Observed changes in molecular structure and organization occurred on the time scale of weeks during sample storage at room temperature. We observed structural changes on faster time scales by heating samples above room temperature or by addition of water. Analysis of hydration effects indicates that water can enhance the ability of the peptide to convert to beta-strand secondary structure and assemble into beta-sheets. However, temperature dependent FTIR and time dependent WAXD data indicate that bound water may hinder the assembly of beta-strands into beta-sheets. We suggest that secondary structural transformation and intermolecular organization together produce a water-insoluble state. These results reveal insights into the role of water in self-assembly of polypeptides with hydrophilic side chains, and have implications on future optimization of RADA16-I nanofiber production.