Design and production of a chimeric resilin-, elastin-, and collagen-like engineered polypeptide.

Protein-inspired biomaterials have gained great interest as an alternative to synthetic polymers, in particular, for their potential use as biomedical devices. The potential inspiring models are mainly proteins able to confer mechanical properties to tissues and organs, such as elasticity (elastin, resilin, spider silk) and strength (collagen, silk). The proper combination of repetitive sequences, each of them derived from different proteins, represents a useful tool for obtaining biomaterials with tailored mechanical properties and biological functions. In this report we describe the design, the production, and the preliminary characterization of a chimeric polypeptide, based on sequences derived from the highly resilient proteins resilin and elastin and from collagen-like sequences. The results show that the obtained chimeric recombinant material exhibits promising self-assembling properties. Young's modulus of the fibers was determined by AFM image analysis and lies in the range of 0.1-3 MPa in agreement with the expectations for elastin-like and resilin-like materials.

[1]  D. Kaplan,et al.  Construction, cloning, and expression of synthetic genes encoding spider dragline silk. , 1995, Biochemistry.

[2]  Xiaoyi Wu,et al.  Recombinant silk-elastinlike protein polymer displays elasticity comparable to elastin. , 2009, Biomacromolecules.

[3]  David A Tirrell,et al.  Physical properties of artificial extracellular matrix protein films prepared by isocyanate crosslinking. , 2004, Biomaterials.

[4]  D. Dudek,et al.  Designed biomaterials to mimic the mechanical properties of muscles , 2010, Nature.

[5]  Alexander K. Buell,et al.  Nanostructured films from hierarchical self-assembly of amyloidogenic proteins. , 2010, Nature nanotechnology.

[6]  J. Castle,et al.  AFM study of the elastin-like biopolymer poly(ValGlyGlyValGly). , 2004, Biomacromolecules.

[7]  Sergiy Minko,et al.  AFM single molecule experiments at the solid-liquid interface: in situ conformation of adsorbed flexible polyelectrolyte chains. , 2005, Journal of the American Chemical Society.

[8]  D. Martin,et al.  Morphology and primary crystal structure of a silk‐like protein polymer synthesized by genetically engineered Escherichia coli bacteria , 1994, Biopolymers.

[9]  Daniel J. Muller,et al.  Observing growth steps of collagen self-assembly by time-lapse high-resolution atomic force microscopy. , 2006, Journal of structural biology.

[10]  Brigida Bochicchio,et al.  Molecular and Supramolecular Structural Studies on Significant Repetitive Sequences of Resilin , 2009, Chembiochem : a European journal of chemical biology.

[11]  S. O. Andersen,et al.  Tentative identification of a resilin gene in Drosophila melanogaster. , 2001, Insect biochemistry and molecular biology.

[12]  Brigida Bochicchio,et al.  Molecular and supramolecular structural studies on human tropoelastin sequences. , 2007, Biophysical journal.

[13]  Y. Popineau,et al.  Expression of a new chimeric protein with a highly repeated sequence in tobacco cells , 2011, Plant Cell Reports.

[14]  Roberto Cingolani,et al.  Amyloid-like fibrils in elastin-related polypeptides: structural characterization and elastic properties. , 2008, Biomacromolecules.

[15]  T. Vuocolo,et al.  Synthesis and properties of crosslinked recombinant pro-resilin , 2005, Nature.

[16]  A. Tamburro,et al.  Elastin-based biopolymers: chemical synthesis and structural characterization of linear and cross-linked poly(OrnGlyGlyOrnGly). , 2002, Biomacromolecules.

[17]  J. Gosline,et al.  Elastic proteins: biological roles and mechanical properties. , 2002, Philosophical transactions of the Royal Society of London. Series B, Biological sciences.

[18]  Brigida Bochicchio,et al.  Investigating by CD the molecular mechanism of elasticity of elastomeric proteins. , 2008, Chirality.

[19]  H. Bennet-Clark THE FIRST DESCRIPTION OF RESILIN , 2007, Journal of Experimental Biology.

[20]  Jason A Burdick,et al.  Hydrophilic elastomeric biomaterials based on resilin-like polypeptides. , 2009, Soft matter.

[21]  J. Rodríguez‐Cabello,et al.  Tailored recombinant elastin-like polymers for advanced biomedical and nano(bio)technological applications , 2006, Biotechnology Letters.

[22]  David L Kaplan,et al.  RGD-functionalized bioengineered spider dragline silk biomaterial. , 2006, Biomacromolecules.

[23]  S. Heilshorn,et al.  Protein-engineered biomaterials: highly tunable tissue engineering scaffolds. , 2010, Tissue engineering. Part B, Reviews.

[24]  T. Asakura,et al.  Synthesis and structural characterization of silk-like materials incorporated with an elastic motif. , 2003, Journal of biochemistry.

[25]  D. Chandler Interfaces and the driving force of hydrophobic assembly , 2005, Nature.

[26]  E. Chaikof,et al.  Viscoelastic and mechanical behavior of recombinant protein elastomers. , 2005, Biomaterials.

[27]  C. Elvin,et al.  Physical approaches for fabrication of organized nanostructure of resilin-mimetic elastic protein rec1-resilin. , 2009, Biomaterials.

[28]  Roberto Cingolani,et al.  Charge transport and intrinsic fluorescence in amyloid-like fibrils , 2007, Proceedings of the National Academy of Sciences.

[29]  J. Castle,et al.  Comment on the mechanical properties of the amyloid fibre, poly(ValGlyGlyLeuGly), obtained by a novel AFM methodology , 2008 .

[30]  A. Pepe,et al.  Exon 26-coded polypeptide: an isolated hydrophobic domain of human tropoelastin able to self-assemble in vitro. , 2008, Matrix biology : journal of the International Society for Matrix Biology.

[31]  A. Bigi,et al.  In vitro calcified tendon collagen: an atomic force and scanning electron microscopy investigation. , 1997, Biomaterials.

[32]  Laurent Kreplak,et al.  Introduction to Atomic Force Microscopy (AFM) in Biology , 2009, Current protocols in protein science.

[33]  F Javier Arias,et al.  Synthesis and characterization of macroporous thermosensitive hydrogels from recombinant elastin-like polymers. , 2009, Biomacromolecules.

[34]  P. L. Atreya,et al.  Interaction of prolyl 4-hydroxylase with synthetic peptide substrates. A conformational model for collagen proline hydroxylation. , 1991, Journal of Biological Chemistry.

[35]  L. Setton,et al.  Rapid cross-linking of elastin-like polypeptides with (hydroxymethyl)phosphines in aqueous solution. , 2007, Biomacromolecules.

[36]  Nobuhiko Saitô,et al.  The Statistical Mechanical Theory of Stiff Chains , 1967 .

[37]  J. Castle,et al.  Transformation of amyloid-like fibers, formed from an elastin-based biopolymer, into a hydrogel: an X-ray photoelectron spectroscopy and atomic force microscopy study. , 2007, Biomacromolecules.

[38]  D. Tirrell,et al.  Engineering the extracellular matrix: a novel approach to polymeric biomaterials. I. Control of the physical properties of artificial protein matrices designed to support adhesion of vascular endothelial cells. , 2000, Biomacromolecules.

[39]  P. Perfetti SCANNING PROBE MICROSCOPY APPLIED TO MATERIALS SCIENCE AND BIOLOGY , 2000 .

[40]  T. Yamaoka,et al.  Mechanism for the phase transition of a genetically engineered elastin model peptide (VPGIG)40 in aqueous solution. , 2003, Biomacromolecules.

[41]  H. Ghandehari,et al.  Genetically engineered silk-elastinlike protein polymers for controlled drug delivery. , 2002, Advanced drug delivery reviews.

[42]  David A Tirrell,et al.  Protein engineering approaches to biomaterials design. , 2005, Current opinion in biotechnology.

[43]  Steven G Wise,et al.  Elastin-based materials. , 2010, Chemical Society reviews.

[44]  G. Fasman,et al.  The evaluation of type I and type II beta-turn mixtures. Circular dichroism, NMR and molecular dynamics studies. , 2009, International journal of peptide and protein research.

[45]  J. G. Vinter,et al.  Thrombogenic collagen-mimetic peptides: Self-assembly of triple helix-based fibrils driven by hydrophobic interactions , 2008, Proceedings of the National Academy of Sciences.

[46]  A. Tamburro,et al.  Chemical synthesis of cross-linked poly(KGGVG), an elastin-like biopolymer. , 2001, Biopolymers.

[47]  Ashutosh Chilkoti,et al.  Stimulus responsive elastin biopolymers: Applications in medicine and biotechnology. , 2006, Current opinion in chemical biology.

[48]  Ashutosh Chilkoti,et al.  Genetically encoded synthesis of protein-based polymers with precisely specified molecular weight and sequence by recursive directional ligation: examples from the elastin-like polypeptide system. , 2002, Biomacromolecules.

[49]  Ashutosh Chilkoti,et al.  Characterization of a genetically engineered elastin-like polypeptide for cartilaginous tissue repair. , 2002, Biomacromolecules.

[50]  A. Fahmi,et al.  Nanopore formation by self-assembly of the model genetically engineered elastin-like polymer [(VPGVG)2(VPGEG)(VPGVG)2]15. , 2004, Journal of the American Chemical Society.

[51]  C. Morrow,et al.  Production and Purification of a Recombinant Elastomeric Polypeptide, G‐(VPGVG)19‐VPGV, from Escherichia coli , 1992, Biotechnology progress.

[52]  David L Kaplan,et al.  Biomaterials derived from silk-tropoelastin protein systems. , 2010, Biomaterials.

[53]  M. Marquet,et al.  Genetic Engineering of Structural Protein Polymers , 1990, Biotechnology progress.

[54]  J. Rodríguez‐Cabello,et al.  Design and bioproduction of a recombinant multi(bio)functional elastin-like protein polymer containing cell adhesion sequences for tissue engineering purposes , 2004, Journal of materials science. Materials in medicine.

[55]  T. Yamaoka,et al.  Effects of temperature and pressure on the aggregation properties of an engineered elastin model polypeptide in aqueous solution. , 2000, Biomacromolecules.

[56]  A. Tamburro,et al.  Synthesis and structural characterization of poly(LGGVG), an elastin-like polypeptide. , 2000, International journal of biological macromolecules.

[57]  Antonio Mario Tamburro,et al.  Biopolymers and biomaterials based on elastomeric proteins , 2002 .

[58]  D. Simionescu,et al.  Elastin stabilization in cardiovascular implants: improved resistance to enzymatic degradation by treatment with tannic acid. , 2004, Biomaterials.

[59]  Chikako Tanaka,et al.  Production and characterization of a silk-like hybrid protein, based on the polyalanine region of Samia cynthia ricini silk fibroin and a cell adhesive region derived from fibronectin. , 2004, Biomaterials.