title: Synthesis of bioactive protein hydrogels by genetically encoded SpyTag-SpyCatcher chemistry.
abstract: 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.
authors: Division of Chemistry and Chemical Engineering, California Institute of Technology, Pasadena, CA 91125.
authors: Fei
authors: F
authors: Sun
authors: Division of Chemistry and Chemical Engineering, California Institute of Technology, Pasadena, CA 91125.
authors: Wen-Bin
authors: WB
authors: Zhang
authors: Division of Chemistry and Chemical Engineering, California Institute of Technology, Pasadena, CA 91125.
authors: Alborz
authors: A
authors: Mahdavi
authors: Division of Chemistry and Chemical Engineering, California Institute of Technology, Pasadena, CA 91125 tirrell@caltech.edu frances@cheme.caltech.edu.
authors: Frances H
authors: FH
authors: Arnold
authors: Division of Chemistry and Chemical Engineering, California Institute of Technology, Pasadena, CA 91125 tirrell@caltech.edu frances@cheme.caltech.edu.
authors: David A
authors: DA
authors: Tirrell
mesh_terms: D016475:3T3 Cells; D000595:Amino Acid Sequence; D000818:Animals; D018914:Cells, Immobilized / Q000166:cytology / Q000187:drug effects / Q000378:metabolism; D053595:Embryonic Stem Cells / Q000166:cytology / Q000187:drug effects / Q000378:metabolism; D005347:Fibroblasts / Q000166:cytology / Q000187:drug effects / Q000378:metabolism; D020100:Hydrogels / Q000494:pharmacology*; D053605:Leukemia Inhibitory Factor / Q000494:pharmacology*; D008164:Luminescent Proteins / Q000378:metabolism; D051379:Mice; D008969:Molecular Sequence Data; D039904:Pluripotent Stem Cells / Q000166:cytology / Q000187:drug effects / Q000378:metabolism; D015202:Protein Engineering / Q000379:methods*; D000097573:Red Fluorescent Protein
chemical_list: D020100:Hydrogels; D053605:Leukemia Inhibitory Factor; D008164:Luminescent Proteins; D000097573:Red Fluorescent Protein
keywords: cell fate control; protein biomaterials; stem cell encapsulation