Synthesis of bioactive protein hydrogels by genetically encoded SpyTag-SpyCatcher chemistry

Protein-based hydrogels have emerged as promising alternatives to synthetic hydrogels for biomedical applications, owing to the precise control of structure and function enabled by protein engineering. Nevertheless, strategies for assembling 3D molecular networks that carry the biological informatio...

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Bibliographic Details
Published in:Proceedings of the National Academy of Sciences - PNAS 2014-08, Vol.111 (31), p.11269-11274
Main Authors: Sun, Fei, Zhang, Wen-Bin, Mahdavi, Alborz, Arnold, Frances H., Tirrell, David A.
Format: Article
Language:English
Subjects:
3T3 Cells
Amino Acid Sequence
Animals
Biological Sciences
Cell adhesion & migration
Cell culture techniques
Cells, Immobilized - cytology
Cells, Immobilized - drug effects
Cells, Immobilized - metabolism
Embryonic Stem Cells - cytology
Embryonic Stem Cells - drug effects
Embryonic Stem Cells - metabolism
Encapsulation
Fibroblasts - cytology
Fibroblasts - drug effects
Fibroblasts - metabolism
Gelation
Gels
Hydrogels
Hydrogels - pharmacology
Leukemia Inhibitory Factor - pharmacology
Luminescent Proteins - metabolism
Mice
Molecular Sequence Data
Peptides
Physical Sciences
Pluripotent stem cells
Pluripotent Stem Cells - cytology
Pluripotent Stem Cells - drug effects
Pluripotent Stem Cells - metabolism
Protein Engineering - methods
Proteins
Red Fluorescent Protein
Stem cells
Teeth
Tissue engineering
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Summary:Protein-based hydrogels have emerged as promising alternatives to synthetic hydrogels for biomedical applications, owing to the precise control of structure and function enabled by protein engineering. Nevertheless, strategies for assembling 3D molecular networks that carry the biological information encoded in full-length proteins remain underdeveloped. Here we present a robust protein gelation strategy based on a pair of genetically encoded reactive partners, SpyTag and SpyCatcher, that spontaneously form covalent isopeptide linkages under physiological conditions. The resulting “network of Spies” may be designed to include cell-adhesion ligands, matrix metalloproteinase-1 cleavage sites, and full-length globular proteins [mCherry and leukemia inhibitory factor (LIF)]. The LIF network was used to encapsulate mouse embryonic stem cells; the encapsulated cells remained pluripotent in the absence of added LIF. These results illustrate a versatile strategy for the creation of information-rich biomaterials.
ISSN:0027-8424
1091-6490
DOI:10.1073/pnas.1401291111