Installing folded proteins into biomaterials is gaining interest for imparting functional properties that often cannot be provided by unfolded peptides or small molecules, such as catalysis, antigen conformation, or molecular recognition. Although covalent grafting provides a simple means to immobilize proteins onto pre-formed biomaterials, amenable chemistries can alter protein bioactivity, are relatively non-specific, and can be difficult to reproduce. Covalent fusions of bioactive molecules and synthetic peptides that can self-assemble into nano-scale architectures are a promising alternative for creating functional supramolecular biomaterials with precise and reproducible composition. Here we created a pair of oppositely charged synthetic peptides, referred to as “CATCH” (Co-Assembly Tags based on CHarge complementarity), to install folded proteins into supramolecular biomaterials. CATCH peptides co-assemble into β-sheet nanofibers when combined, yet cannot assemble independently due to electrostatic repulsion. Electrostatically controlled assembly enabled high yield production of soluble CATCH-green fluorescent protein (CATCH(−)GFP) by E. coli. Binary mixtures of CATCH-GFP and its charge-complementary peptide self-assembled into fluorescent microparticles, whereas ternary mixtures of CATCH(−)GFP and both CATCH peptides self-assembled into fluorescent nanofibers and macroscopic hydrogels. The CATCH system is therefore likely to be broadly useful for creating functional supramolecular biomaterials with integrated folded protein components for various biomedical and biotechnological applications.
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This work was supported by start-up funds from the University of Florida Office of the Provost, the Herbert Wertheim College of Engineering, and the J. Crayton Pruitt Family Department of Biomedical Engineering. Anthony D. Sorrentino was supported by a fellowship from the University of Florida University Scholars Program.
D.T.S designed and conducted experiments, analyzed data, and wrote the paper. A.R. assisted with TEM. A.D.S. assisted with SEC. K.R.K. assisted with peptide synthesis and purification. S.J.H. assisted with CD, analyzed the results, and edited the paper. G.A.H. directed the work, designed experiments, analyzed results, and wrote the paper.
Conflict of interest
The authors, Dillon T. Seroski, Antonietta Restuccia, Anthony D. Sorrentino, Kevin R. Knox, Stephen J. Hagen, and Gregory A. Hudalla, declare no conflicts of interest.
No human studies or animal studies were carried out by the authors for this article.
Associate Editor Michael R. King oversaw the review of this article.
Gregory A. Hudalla is an Assistant Professor in the J. Crayton Pruitt Family Department of Biomedical Engineering in the Wertheim College of Engineering at the University of Florida. He received his B.S. in Chemical Engineering from the Illinois Institute of Technology, and received his M.S. and Ph.D. in Biomedical Engineering from the University of Wisconsin, under the guidance of Dr. William L. Murphy. Prior to joining the University of Florida, Dr. Hudalla was a postdoctoral scholar at the University of Chicago and Northwestern University under the guidance of Dr. Joel H. Collier and Dr. Milan Mrksich. Research in the Hudalla lab creates functional biomaterials for therapeutic or diagnostic applications via “self-assembly”, the spontaneous organization of molecules into supramolecular architectures. A primary research emphasis is developing synthetic peptide “fusion tags” to organize bioactive molecules, such as carbohydrates and proteins, into functional supramolecular materials. Dr. Hudalla is the recipient of a 2015 NSF Career award in Biomaterials and is supported by research grants from the NIH.
This article is part of the 2016 Young Innovators Issue.
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Seroski, D.T., Restuccia, A., Sorrentino, A.D. et al. Co-Assembly Tags Based on Charge Complementarity (CATCH) for Installing Functional Protein Ligands into Supramolecular Biomaterials. Cel. Mol. Bioeng. 9, 335–350 (2016). https://doi.org/10.1007/s12195-016-0459-2