title: Tandem modular protein-based hydrogels constructed using a novel two-component approach.
abstract: Leucine zipper sequences have been widely used to engineer protein-based hydrogels for biomedical applications. Previously, we have used this method to engineer tandem modular protein-based hydrogels as a step toward developing extracellular matrix-mimetic hydrogels. However, the spontaneous self-association of leucine zipper sequences in solution has made it challenging to express and purify tandem modular proteins carrying leucine zipper under native conditions. To obviate this problem, here we report a novel two-component approach to engineer tandem modular protein-based hydrogels. This methodology makes use of two complementary leucine zipper sequences (CCE and CCK), which do not self-associate but self-assemble into heterodimeric coiled-coils at neutral pH, as functional groups to drive the self-assembly of protein hydrogels. The two protein components are bifunctional and trifunctional tandem modular proteins carrying the leucine zipper functional groups. We found that the two proteins carrying CCE or CCK can be expressed and purified under native conditions with high yield. Upon mixing, the aqueous solution of the two proteins readily forms a transparent hydrogel. The resultant hydrogel can undergo reversible sol-gel transitions as a function of temperature, and shows much improved erosion properties. This method provides a new approach to tune the topology and physical properties of the protein hydrogels via genetic engineering, and opens the possibility to systematically explore the use of large native extracellular proteins to engineer extracellular matrix-mimetic hydrogels.
authors.affiliation: Department of Chemistry, University of British Columbia, Vancouver, BC V6T 1Z1 Canada.
authors.forename: Shanshan
authors.initials: S
authors.lastname: Lv
authors.forename: Yi
authors.initials: Y
authors.lastname: Cao
authors.forename: Hongbin
authors.initials: H
authors.lastname: Li
mesh_terms: D000595:Amino Acid Sequence; D004738:Engineering / Q000379:methods*; D005612:Freeze Drying; D020100:Hydrogels / Q000737:chemistry*; D016350:Leucine Zippers; D008969:Molecular Sequence Data; D010446:Peptide Fragments / Q000737:chemistry; D017434:Protein Structure, Tertiary; D011506:Proteins / Q000737:chemistry*; D014867:Water / Q000737:chemistry
chemical_list: D020100:Hydrogels; D010446:Peptide Fragments; D011506:Proteins; D014867:Water