title: 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.
authors.affiliation: School of Chemical Engineering, Purdue University , West Lafayette, Indiana 47907-2100, United States.
authors.forename: Renay S-C
authors.initials: RS
authors.lastname: Su
authors.forename: Julie N
authors.initials: JN
authors.lastname: Renner
authors.forename: Julie C
authors.initials: JC
authors.lastname: Liu
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D002448:Cell Adhesion; D002465:Cell Movement; D002470:Cell Survival / Q000187:drug effects; D002478:Cells, Cultured; D020099:Coated Materials, Biocompatible / Q000737:chemistry; D004926:Escherichia coli; D061307:Human Umbilical Vein Endothelial Cells / Q000502:physiology; D006801:Humans; D008422:Materials Testing; D049897:Pectinidae; D010446:Peptide Fragments / Q000096:biosynthesis / Q000737:chemistry* / Q000633:toxicity; D017433:Protein Structure, Secondary; D011506:Proteins / Q000737:chemistry*; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000737:chemistry / Q000633:toxicity; D013499:Surface Properties
chemical_list: C023706:Abductin; D020099:Coated Materials, Biocompatible; D010446:Peptide Fragments; D011506:Proteins; D011994:Recombinant Proteins