The overall objective of this competing R01 renewal is the continued development of the Elastin Fusion Protein technology as a genetically encodable, bioinspired analog of "smart" -stimulus responsive- protein-polymer conjugates. In this renewal application, we seek to greatly expand upon our ability to modulate the solution behavior of ELP fusion proteins by introducing a greater level of sophistication beyond simple physical triggers of the ELP phase transition and uncontrolled aggregation as the final state, by developing new strategies for the: (1) stimulus responsive self-assembly of ELP fusion proteins into micelles or other self-assembled nano-mesoscale structures;and (2) coupling molecular recognition to the phase transition of ELPs, so that ELP fusion proteins can be programmed to exhibit an isothermal phase transition in response to ligand-binding. These "second-generation" ELP fusion proteins will, we believe, provide a new set of applications in Biotechnology and Medicine by virtue of the increased sophistication of the functionality embedded in these ELPs. For example, the ability to trigger an LCST transition by biochemical ligand-binding to an ELP fusion protein will provide a simple, convenient and modular approach to design biopolymer actuators that exhibit ligand-binding triggered phase transition behavior by splicing together ligand binding domains with ELPs. Similarly, developing a generic methodology to convert a peptide or protein into a nanoscale construct such as a spherical micelle has implications for the delivery of protein therapeutics because: (1) it will increase the apparent size of the biomolecule and will thereby increase its in vivo half-life;and (2) will provide a rational strategy to enhance the affinity and specificity of a protein or peptide drug for its target receptor via multivalency.

Public Health Relevance

This project will develop bioinspired "smart" -stimulus responsive- Elastin fusion proteins. In this renewal application, we seek to greatly expand upon our ability to modulate the solution behavior of ELP fusion proteins by developing a new generation of ELP fusion proteins that will self-assemble into nanoparticles of defined size and shape or that will exhibit soluble-insoluble phase transition behavior upon binding to another molecule. These ELP fusion proteins will provide new molecular building blocks that will enable new approaches for targeted drug delivery, regenerative medicine and bioanalytical assays.

National Institute of Health (NIH)
National Institute of General Medical Sciences (NIGMS)
Research Project (R01)
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Biomaterials and Biointerfaces Study Section (BMBI)
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Smith, Ward
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Duke University
Biomedical Engineering
Schools of Engineering
United States
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Bellucci, Joseph J; Bhattacharyya, Jayanta; Chilkoti, Ashutosh (2015) A noncanonical function of sortase enables site-specific conjugation of small molecules to lysine residues in proteins. Angew Chem Int Ed Engl 54:441-5
McDaniel, Jonathan R; Weitzhandler, Isaac; Prevost, Sylvain et al. (2014) Noncanonical self-assembly of highly asymmetric genetically encoded polypeptide amphiphiles into cylindrical micelles. Nano Lett 14:6590-8
Hassouneh, Wafa; Nunalee, Michelle L; Shelton, M Coleman et al. (2013) Calcium binding peptide motifs from calmodulin confer divalent ion selectivity to elastin-like polypeptides. Biomacromolecules 14:2347-53
Thapa, Arjun; Han, Wei; Simons, Robin H et al. (2013) Effect of detergents on the thermal behavior of elastin-like polypeptides. Biopolymers 99:55-62
Christensen, Trine; Hassouneh, Wafa; Trabbic-Carlson, Kimberley et al. (2013) Predicting transition temperatures of elastin-like polypeptide fusion proteins. Biomacromolecules 14:1514-9
Kurzbach, Dennis; Hassouneh, Wafa; McDaniel, Jonathan R et al. (2013) Hydration layer coupling and cooperativity in phase behavior of stimulus responsive peptide polymers. J Am Chem Soc 135:11299-308
Hassouneh, Wafa; MacEwan, Sarah R; Chilkoti, Ashutosh (2012) Fusions of elastin-like polypeptides to pharmaceutical proteins. Methods Enzymol 502:215-37
Asai, Daisuke; Xu, Donghua; Liu, Wenge et al. (2012) Protein polymer hydrogels by in situ, rapid and reversible self-gelation. Biomaterials 33:5451-8
Hassouneh, Wafa; Fischer, Karl; MacEwan, Sarah R et al. (2012) Unexpected multivalent display of proteins by temperature triggered self-assembly of elastin-like polypeptide block copolymers. Biomacromolecules 13:1598-605
Amiram, Miriam; Quiroz, Felipe Garcia; Callahan, Daniel J et al. (2011) A highly parallel method for synthesizing DNA repeats enables the discovery of 'smart' protein polymers. Nat Mater 10:141-8

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