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Using Tools from Optogenetics to Create Light-Responsive Biomaterials: LOVTRAP-PEG Hydrogels for Dynamic Peptide Immobilization

  • S.I. : Biomaterials - Engineering Cell Behavior
  • Published:
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Abstract

Hydrogel materials have become a versatile platform for in vitro cell culture due to their ability to simulate many aspects of native tissues. However, precise spatiotemporal presentation of peptides and other biomolecules has remained challenging. Here we report the use of light-sensing proteins (LSPs), more commonly used in optogenetics research, as light-activated reversible binding sites within synthetic poly(ethylene glycol) (PEG) hydrogels. We used LOVTRAP, a two component LSP system consisting of LOV2, a protein domain that can cycle reversibly between “light” and “dark” conformations in response to blue light, and a z-affibody, Zdark (Zdk), that binds the dark state of LOV2, to spatiotemporally control the presentation of a recombinant protein within PEG hydrogels. By immobilizing LOV2 within PEG gels, we were able to capture a recombinant fluorescent protein (used as a model biomolecule) containing a Zdk domain, and then release the Zdk fusion protein using blue light. Zdk was removed from LOV2-containing PEG gels using focused blue light, resulting in a 30% reduction of fluorescence compared to unexposed regions of the gel. Additionally, the reversible binding capability of LOVTRAP was observed in our system, enabling our LOV2 gels to capture and release Zdk at least three times. By adding a Zdk domain to a recombinant peptide or protein, dynamic, spatially constrained displays of non-diffusing ligands within a PEG gel could feasibly be achieved using LOV2.

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Acknowledgments

We would like to acknowledge the generosity of Dr. Charles Gersbach for providing the blue LED array used in this study and Dr. Ashutosh Chilkoti for providing the sortase-ELP construct used to conjugate LOV2 to PEG. This work was supported by the National Science Foundation Graduate Research Fellowship under Grant Number DGE1644868.

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The authors have no conflicts of interest to report.

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  1. Department of Biomedical Engineering, Duke University, 101 Science Drive Campus Box 90281, Durham, NC, 27708-0281, USA

    Joshua A. Hammer, Anna Ruta & Jennifer L. West

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Correspondence to Jennifer L. West.

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Hammer, J.A., Ruta, A. & West, J.L. Using Tools from Optogenetics to Create Light-Responsive Biomaterials: LOVTRAP-PEG Hydrogels for Dynamic Peptide Immobilization. Ann Biomed Eng 48, 1885–1894 (2020). https://doi.org/10.1007/s10439-019-02407-w

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