Abstract
Rec1-resilin is the first recombinant resilin-mimetic protein polymer, synthesized from exon-1 of the Drosophila melanogaster gene CG15920 that has demonstrated unusual multi-stimuli responsiveness in aqueous solution. Crosslinked hydrogels of Rec1-resilin have also displayed remarkable mechanical properties including near-perfect rubber-like elasticity. The structural basis of these extraordinary properties is not clearly understood. Here we combine a computational and experimental investigation to examine structural ensembles of Rec1-resilin in aqueous solution. The structure of Rec1-resilin in aqueous solutions is investigated experimentally using circular dichroism (CD) spectroscopy and small angle X-ray scattering (SAXS). Both bench-top and synchrotron SAXS are employed to extract structural data sets of Rec1-resilin and to confirm their validity. Computational approaches have been applied to these experimental data sets in order to extract quantitative information about structural ensembles including radius of gyration, pair-distance distribution function, and the fractal dimension. The present work confirms that Rec1-resilin is an intrinsically disordered protein (IDP) that displays equilibrium structural qualities between those of a structured globular protein and a denatured protein. The ensemble optimization method (EOM) analysis reveals a single conformational population with partial compactness. This work provides new insight into the structural ensembles of Rec1-resilin in solution.
Highlights
Million cycles), and its role in the jumping, flying and sound production mechanisms in insects[11]
We present a comprehensive description of the equilibrium structural ensembles for Rec1-resilin, which underpins the molecular basis of its properties
The secondary structure of Rec1-resilin was predicted from its primary amino acid sequence (Fig. 1 in Supplementary Information) using different key secondary structure modelling routines including DSC (Discrimination of protein Secondary structure Class)[30], PHDsec (Profile network prediction HeiDelberg secondary structure)[31], and SOPMA (Self-Optimized Prediction Method with Alignment)[32]
Summary
Million cycles), and its role in the jumping, flying and sound production mechanisms in insects[11]. Owing to the low stiffness, high fatigue lifetime, resilience, and attractive biochemical and mechanical properties, cross-linked RMPs have emerged as valuable materials for biomedical applications including tissue engineering[3,14,15,16,17]. Rec1-resilin exhibits multi-stimuli responsiveness including thermal, pH, ion, and photo responsiveness in aqueous solution[19]. It displays unusual dual phase thermal transition behaviour, i.e. upper critical solution temperature (UCST) as well as lower critical solution temperature (LCST)[19]. Cross-linked Rec1-resilin hydrogels exhibit near-perfect rubber-like elasticity with outstanding resilience (> 92%) and negligible creep behaviour[18,24]. The molecular and structural basis of the characteristics of Rec1-resilin in solution and of the cross-linked Rec1-resilin hydrogels has not yet been fully elucidated. We present a comprehensive description of the equilibrium structural ensembles for Rec1-resilin, which underpins the molecular basis of its properties
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