title: Synthesis of an intein-mediated artificial protein hydrogel.
abstract: We present the synthesis of a highly stable protein hydrogel mediated by a split-intein-catalyzed protein trans-splicing reaction. The building blocks of this hydrogel are two protein block-copolymers each containing a subunit of a trimeric protein that serves as a crosslinker and one half of a split intein. A highly hydrophilic random coil is inserted into one of the block-copolymers for water retention. Mixing of the two protein block copolymers triggers an intein trans-splicing reaction, yielding a polypeptide unit with crosslinkers at either end that rapidly self-assembles into a hydrogel. This hydrogel is very stable under both acidic and basic conditions, at temperatures up to 50 °C, and in organic solvents. The hydrogel rapidly reforms after shear-induced rupture. Incorporation of a "docking station peptide" into the hydrogel building block enables convenient incorporation of "docking protein"-tagged target proteins. The hydrogel is compatible with tissue culture growth media, supports the diffusion of 20 kDa molecules, and enables the immobilization of bioactive globular proteins. The application of the intein-mediated protein hydrogel as an organic-solvent-compatible biocatalyst was demonstrated by encapsulating the horseradish peroxidase enzyme and corroborating its activity.
authors: Artie McFerrin Department of Chemical Engineering, Texas A&M University, College Station.
authors: Miguel A
authors: MA
authors: Ramirez
authors: Zhilei
authors: Z
authors: Chen
mesh_terms: D004800:Enzymes, Immobilized / Q000737:chemistry; D049452:Green Fluorescent Proteins / Q000737:chemistry; D006735:Horseradish Peroxidase / Q000737:chemistry; D020100:Hydrogels / Q000138:chemical synthesis*; D006863:Hydrogen-Ion Concentration; D055767:Immobilized Proteins / Q000737:chemistry; D047668:Inteins; D055550:Protein Stability; D011506:Proteins / Q000138:chemical synthesis*
chemical_list: D004800:Enzymes, Immobilized; D020100:Hydrogels; D055767:Immobilized Proteins; D011506:Proteins; D049452:Green Fluorescent Proteins; D006735:Horseradish Peroxidase