Multi-scale characterization of thermoresponsive dendritic elastin-like peptides

Elastin like peptides (ELPs)-polypeptides based on the protein elastin-are used widely as thermoresponsive components in biomaterials due to the presence of a sharp soluble-to-insoluble phase change at a characteristic transition temperature (T_t). While linear ELPs have been thoroughly studied, few investigations into branched ELPs have been carried out. Using lysine amino acids as branching and terminal units with 1-3 pentameric repeats between each branch, ELP dendrimers were prepared by solid-phase peptide synthesis with molecular weights as high as 14kDa. A conformation change from random coil to β-turn upon heating through the T_t, typical of ELPs, was observed by circular dichroism spectroscopy for all peptides. The high molecular weights of these peptides enabled the use of characterization techniques typically reserved for polymers. Variable-temperature small-angle X-ray scattering measurements in dilute solution revealed an increase in size and fractal dimension upon heating, even well below the T_t. These results were corroborated by cryogenic transmission electron microscopy, which confirmed the presence of aggregates below the T_t, and micro differential scanning calorimetry, which showed a broad endothermic peak below the T_t. These results collectively indicate the presence of a pre-coacervation step in the phase transition of ELP dendrimers.