title: Engineering of an environmentally responsive beta roll peptide for use as a calcium-dependent cross-linking domain for peptide hydrogel formation.
abstract: We have created a set of rationally designed peptides that form calcium-dependent hydrogels based on the beta roll peptide domain. In the absence of calcium, the beta roll domain is intrinsically disordered. Upon the addition of calcium, the peptide forms a beta helix secondary structure. We have designed two variations of our beta roll domain. First, we have mutated one face of the beta roll domain to contain leucine residues so that the calcium-dependent structural formation leads to dimerization through hydrophobic interactions. Second, an α-helical leucine zipper domain is appended to the engineered beta roll domain as an additional means of forming intermolecular cross-links. This full peptide construct forms a hydrogel only in calcium-rich environments. The resulting structural and mechanical properties of the supramolecular assemblies are compared with the wild-type domain using several biophysical techniques including circular dichroism, FRET, bis-ANS binding and microrheology. The calcium responsiveness and rheological properties of the leucine beta roll containing construct confirm the potential of this allosterically regulated scaffold to serve as a cross-linking domain for stimulus-responsive biomaterials development.
authors.affiliation: Department of Chemical Engineering, Columbia University, New York, New York, United States.
authors.forename: Kevin
authors.initials: K
authors.lastname: Dooley
authors.forename: Yang Hee
authors.initials: YH
authors.lastname: Kim
authors.forename: Hoang D
authors.initials: HD
authors.lastname: Lu
authors.forename: Raymond
authors.initials: R
authors.lastname: Tu
authors.forename: Scott
authors.initials: S
authors.lastname: Banta
mesh_terms: D002118:Calcium / Q000737:chemistry*; D003432:Cross-Linking Reagents / Q000138:chemical synthesis* / Q000737:chemistry; D020136:Hydrogel, Polyethylene Glycol Dimethacrylate / Q000138:chemical synthesis* / Q000737:chemistry; D007930:Leucine / Q000737:chemistry*; D046911:Macromolecular Substances / Q000138:chemical synthesis / Q000737:chemistry; D008958:Models, Molecular; D010455:Peptides / Q000138:chemical synthesis* / Q000737:chemistry; D015202:Protein Engineering; D017433:Protein Structure, Secondary
chemical_list: D003432:Cross-Linking Reagents; D046911:Macromolecular Substances; D010455:Peptides; D020136:Hydrogel, Polyethylene Glycol Dimethacrylate; D007930:Leucine; D002118:Calcium