Synthesis and Characterization of Recombinant Abductin-Based Proteins

Abstract

Recombinant proteins are promising tools for tissue engineering and drug delivery applications. Protein-based biomaterials have several advantages over natural and synthetic polymers, including precise control over amino acid composition and molecular weight, modular swapping of functional domains, and tunable mechanical and physical properties. In this work, we describe recombinant proteins based on abductin, an elastomeric protein that is found in the inner hinge of bivalves and functions as a coil spring to keep shells open. We illustrate, for the first time, the design, cloning, expression, and purification of a recombinant protein based on consensus abductin sequences derived from Argopecten irradians . The molecular weight of the protein was confirmed by mass spectrometry, and the protein was 94% pure. Circular dichroism studies showed that the dominant structures of abductin-based proteins were polyproline II helix structures in aqueous solution and type II β-turns in trifluoroethanol. Dynamic light scattering studies illustrated that the abductin-based proteins exhibit reversible upper critical solution temperature behavior and irreversible aggregation behavior at high temperatures. A LIVE/DEAD assay revealed that human umbilical vein endothelial cells had a viability of 98 ± 4% after being cultured for two days on the abductin-based protein. Initial cell spreading on the abductin-based protein was similar to that on bovine serum albumin. These studies thus demonstrate the potential of abductin-based proteins in tissue engineering and drug delivery applications due to the cytocompatibility and its response to temperature.

Resilin-PEG Hybrid Hydrogels Yield Degradable Elastomeric Scaffolds with Heterogeneous Microstructure

“…19–21 For example, collagen and elastin serve as critical structural components of cardiovascular tissues, which experience cyclic hemodynamic forces over extended time scales, and desirable biomaterials for cardiovascular applications would replicate the elasticity conferred by those proteins. 21–24 Intrinsically disordered proteins (IDP) that serve as structural components of tissue such as elastin, resilin, abductin, and flagelliform spider silk are attractive candidates as these proteins display remarkable elasticity and fatigue resistance; 11, 25–27 protein polymers based upon the characteristic sequences of IDPs have demonstrated mechanical properties comparable to those of the natural proteins. 25 In particular, recently developed resilin-like polypeptide (RLPs) hydrogels have been shown to form elastic materials that are stable, resilient, and highly extensible.…”

Structural ensembles reveal intrinsic disorder for the multi-stimuli responsive bio-mimetic protein Rec1-resilin

“…Recombinant protein synthesis techniques provide an attractive platform for the development of new soft materials with precise control over the composition (monomer sequence), size, structure and functions 5 6 . Several biomimetic protein polymers including variants of green fluorescent protein 7 , elastin 8 , resilin 3 9 and abductin 10 have been reported in the literature highlighting numerous features including responsiveness, organization, and functionality.…”

Phase separation and mechanical properties of an elastomeric biomaterial from spider wrapping silk and elastin block copolymers

“…[50][51][52] In addition to elastin, elastomeric proteins such as silks, resilin, and abductin undergo a liquid-liquid phase separation, where the process is variously suggested to concentrate monomers, control protein-protein interactions to protect from premature aggregation, juxtapose residues for cross-linking as necessary, and be on-pathway for fiber formation. 5,48,53,54 Spinning of silk fibers from a liquid crystalline solution (dope) is tightly regulated to control the stability and solubility of silk proteins, and the formation and alignment of structure. 22 The process involves progressive phase separation and precipitation of the silk as it is drawn from the gland down the duct, with evidence for molten globule states and sol-gel transitions.…”