Functionalization and Patterning of Protein‐Based Materials Using Active Ultrabithorax Chimeras

Zhao, Huayan; Salim, Taha; Brawley, Autumn; Patterson, Jan L.; Matthews, Kathleen S.; Bondos, Sarah E.

doi:10.1002/adfm.201100067

Cited by 16 publications

(77 citation statements)

References 42 publications

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“…19 The plasmid construct was transformed into BL21(DE3)pLysS E. coli cells. E. coli cultures were grown in Luria broth containing 50 μg/mL carbenicillin and 30 μg/mL chloramphenicol (LB) at 37°C.…”

Section: Egfp-ubx Protein Expression and Purificationmentioning

confidence: 99%

On the Design of Composite Protein–Quantum Dot Biomaterials via Self-Assembly

et al. 2011

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Incorporation of nanoparticles during the hierarchical self-assembly of protein-based materials can impart function to the resulting composite materials. Herein we demonstrate that the structure and nanoparticle distribution of composite fibers are sensitive to the method of nanoparticle addition and the physicochemical properties of both the nanoparticle and the protein. Our model system consists of a recombinant enhanced green fluorescent protein-Ultrabithorax (EGFP-Ubx) fusion protein and luminescent CdSe-ZnS core-shell quantum dots (QDs), allowing us to optically assess the distribution of both the protein and nanoparticle components within the composite material. Although QDs favorably interact with EGFP-Ubx monomers, the relatively rough surface morphology of composite fibers suggests EGFP-Ubx-QD conjugates impact self-assembly. Indeed, QDs templated onto EGFP-Ubx film post-self-assembly can be subsequently drawn into smooth composite fibers. Additionally, the QD surface charge impacts QD distribution within the composite material, indicating that surface charge plays an important role in self-assembly. QDs with either positively or negatively charged coatings significantly enhance fiber extensibility. Conversely, QDs coated with hydrophobic moieties and suspended in toluene produce composite fibers with a heterogeneous distribution of QDs and severely altered fiber morphology, indicating that toluene severely disrupts Ubx self-assembly. Understanding factors that impact the protein-nanoparticle interaction enables manipulation of the structure and mechanical properties of composite materials. Since proteins interact with nanoparticle surface coatings, these results should be applicable to other types of nanoparticles with similar chemical groups on the surface.

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Section: Egfp-ubx Protein Expression and Purificationmentioning

confidence: 99%

On the Design of Composite Protein–Quantum Dot Biomaterials via Self-Assembly

et al. 2011

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“…Ubx fibers are compatible with primary human ECs in culture Because Ubx materials have properties that could be useful in tissue engineering studies, 9,11 we tested whether these materials are directly and indirectly compatible with cells in culture.…”

Section: Resultsmentioning

confidence: 99%

Materials composed of theDrosophila melanogasterprotein ultrabithorax are cytocompatible

Patterson

Abbey

Bayless

et al. 2013

J. Biomed. Mater. Res.

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The Drosophila melanogaster Hox protein ultrabithorax (Ubx) has the interesting ability to hierarchically self-assemble in vitro into materials that have mechanical properties comparable to natural elastin. Ubx materials can be easily functionalized by gene fusion, generating potentially useful scaffolds for cell and tissue engineering. Here, we tested the cytocompatibility of fibers composed of Ubx or an mCherry-Ubx fusion protein. Fibers were cultured with three primary human cell lines derived from vasculature at low passage: umbilical vein endothelial cells, brain vascular pericytes, or aortic smooth muscle cells. No direct or indirect toxicity was observed for any cell line, in response to fibers composed of either plain Ubx or mCherry-Ubx. Cells readily adhered to Ubx fibers, and cells attached to fibers could be transferred between tissue cultures without loss of viability for at least 96 h. When attached to fibers, the morphology of the three cell lines differed somewhat, but all cells in contact with Ubx fibers exhibited a microtubular network aligned with the long axis of Ubx fibers. Thus, Ubx fibers are cytocompatible with cultured primary human vascular cells.

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“…Monomers of his‐tagged Ultrabithorax splicing isoform Ia were produced in E. coli from the pET19b‐UbxIa vector and purified as previously described . Ubx fibers were produced by the tray/buffer reservoir system, also as previously described …”

Section: Methodsmentioning

confidence: 99%

“…Protein forms a major component of the extracellular matrix, and protein‐based materials can have chemical and mechanical properties similar to the natural cell environment, which is important for the correct specification of cell behavior . Recombinant protein monomers can also be readily functionalized by gene fusion, in which DNA encoding a functional peptide or protein is fused to DNA encoding the self‐assembling protein . Finally, recombinant production of proteins provides a renewable and uniform supply of monomers for assembly whose sequences, and hence properties, can be easily engineered .…”

Section: Introductionmentioning

confidence: 99%

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Materials composed of theDrosophilaHox protein Ultrabithorax are biocompatible and nonimmunogenic

Patterson

Arenas-Gamboa

Wang

et al. 2014

J. Biomed. Mater. Res.

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Although the in vivo function of the Drosophila melanogaster Hox protein Ultrabithorax (Ubx) is to regulate transcription, in vitro Ubx hierarchically self-assembles to form nanoscale to macroscale materials. The morphology, mechanical properties, and functionality (via protein chimeras) of Ubx materials are all easily engineered. Ubx materials are also compatible with cells in culture. These properties make Ubx attractive as a potential tissue engineering scaffold, but to be used as such they must be biocompatible and nonimmunogenic. In this study, we assess whether Ubx materials are suitable for in vivo applications. When implanted into mice, Ubx fibers attracted few immune cells to the implant area. Sera from mice implanted with Ubx contain little to no antibodies capable of recognizing Ubx. Furthermore, Ubx fibers cultured with macrophages in vitro did not lyse or activate the macrophages, as measured by TNF-α and NO secretion. Finally, Ubx fibers do not cause hemolysis when incubated with human red blood cells. The minimal effects observed are comparable with those induced by biomaterials used successfully in vivo. We conclude Ubx materials are biocompatible and nonimmunogenic.

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Functionalization and Patterning of Protein‐Based Materials Using Active Ultrabithorax Chimeras

Cited by 16 publications

References 42 publications

On the Design of Composite Protein–Quantum Dot Biomaterials via Self-Assembly

On the Design of Composite Protein–Quantum Dot Biomaterials via Self-Assembly

Materials composed of theDrosophila melanogasterprotein ultrabithorax are cytocompatible

Materials composed of theDrosophilaHox protein Ultrabithorax are biocompatible and nonimmunogenic

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