Investigation on the Mechanisms of Elastomechanical Behavior of Resilin
Resilin is a disordered elastomeric protein and can be found in specialized regions of insect cuticles. Its protein sequence, functions and dynamic mechanical properties vary substantially across the species. Resilin can operate across the frequency range from 5 Hz for locomotion to 13 kHz for sound production. To understand the functions of different exons of resilin, we synthesize recombinant resilin-like hydrogels from different exons, and investigate the water content and dynamic mechanical properties, along with estimating surface energies relevant for adhesion. The recombinant resilin-like hydrogel has 80wt% water and does not show any sign of tack even though it satisfies the Dahlquist criterion. Finally, doubly shifted dynamic moduli master curves are developed by applying the time-temperature concentration superposition principle (TTCSP), and compared to results obtained with natural resilin from locusts, dragonflies and cockroaches. The resulting master curves show that the synthetic resilin undergoes a prominent transition, though the responsible mechanism is unclear. Possible explanations for the significant increase in modulus include the formation of intramolecular hydrogen bonds, altered structural organization, or passing through a glass transition, all of which have been reported in the literature for polymeric materials. Results show that in nature, resilin operates at a much lower frequency than this glass transition frequency at room temperature. Moreover, recombinant resilins from different clones have comparable resilience with natural resilin, though the modulus is around 1.5 decades lower. Results from the clones with and without chitin binding domains (ChBD) indicate that the transition for the clone without ChBD occurs at lower frequencies than for those with the ChBD, perhaps due to the disordered nature of the clone without ChBD.
Atomistic molecular modeling is applied on the repetitive motifs of resilin and different elastomeric proteins to better understand the relationship between elastomeric behavior and amino acid sequences. Results show that the motifs form a favorable bent conformation, likely enabled by glycine's lack of steric hindrance and held in place through intramolecular hydrogen bonds. During Steered Molecular Dynamic (SMD) pulling of these motifs, the hydrogen bonds break and they reform again when the peptides are released to move freely, returning to similar bent conformations. The transition seen in the master curves of recombinant resilins might be due to either these intramolecular hydrogen bonds or to glass transition behavior, though evidence indicates that the transition probably due to the glass transition. What we learned from the synthesized recombinant resilin and simulating the repetitive motifs of resilin may be applicable to the biology and mechanics of other elastomeric biomaterials, and may provide deeper understanding of their unique properties.