Ramirez Miguel A, Chen Zhilei
Department of Chemical Engineering, Texas A&M University, College Station, TX, 77843, USA.
Department of Microbial and Molecular Pathogenesis, Texas A&M University Health Science Center, College Station, TX, 77843, USA.
Methods Mol Biol. 2017;1495:161-171. doi: 10.1007/978-1-4939-6451-2_11.
Proteins are nature's building blocks and indispensable in living organisms. Protein-based hydrogels have a wide variety of applications in research and biotechnology. In this chapter, we describe an intein-mediated protein hydrogel that utilizes two synthetic soluble protein block copolymers, each containing a subunit of a trimeric protein that serves as a cross-linker and one half of the naturally split DnaE intein from Nostoc punctiforme. Mixing of these two protein block copolymers initiates an intein trans-splicing reaction that constitutes a self-assembling polypeptide flanked by cross-linkers, triggering protein hydrogel formation. The generated hydrogels are highly stable under both acidic and basic conditions, and at temperatures up to 50 °C. In addition, these hydrogels are able to undergo rapid reassembly after shear-induced rupture. Incorporation of an appropriate binding motif into the protein block copolymers enables the convenient site-specific incorporation of functional globular proteins into the hydrogel network.
