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.
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.
|Nawroth, Jonas F; McDaniel, Jonathan R; Chilkoti, Ashutosh et al. (2016) Maleimide-Functionalized Poly(2-Oxazoline)s and Their Conjugation to Elastin-Like Polypeptides. Macromol Biosci 16:322-33|
|Tang, Nicholas C; Chilkoti, Ashutosh (2016) Combinatorial codon scrambling enables scalable gene synthesis and amplification of repetitiveÂ proteins. Nat Mater 15:419-24|
|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|
|Roberts, Stefan; Dzuricky, Michael; Chilkoti, Ashutosh (2015) Elastin-like polypeptides as models of intrinsically disordered proteins. FEBS Lett 589:2477-86|
|Quiroz, Felipe GarcÃa; Chilkoti, Ashutosh (2015) Sequence heuristics to encode phase behaviour in intrinsically disordered protein polymers. Nat Mater 14:1164-71|
|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|
|Christensen, Trine; Hassouneh, Wafa; Trabbic-Carlson, Kimberley et al. (2013) Predicting transition temperatures of elastin-like polypeptide fusion proteins. Biomacromolecules 14:1514-9|
|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|
|Bellucci, Joseph J; Amiram, Miriam; Bhattacharyya, Jayanta et al. (2013) Three-in-one chromatography-free purification, tag removal, and site-specific modification of recombinant fusion proteins using sortase A and elastin-like polypeptides. Angew Chem Int Ed Engl 52:3703-8|
|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|
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