ABSTRACT
Artificial mussel-glue proteins with pH-triggered cohesion control were synthesized by extending the tyrosinase activated polymerization of peptides to sequences with specific modules for cohesion control. The high propensity of these sequence sections to adopt ß-sheets is suppressed by switch defects. This allows enzymatic activation and polymerization to proceed undisturbed. The ß-sheet formation is regained after polymerization by changing the pH from 5.5 to 6.8, thereby triggering OâN acyl transfer rearrangements that activate the cohesion mechanism. The resulting artificial mussel glue proteins exhibit rapid adsorption on alumina surfaces. The coatings resist harsh hypersaline conditions, and reach remarkable adhesive energies of 2.64â mJ m-2 on silica at pHâ 6.8. In inâ situ switch experiments, the minor pH change increases the adhesive properties of a coating by 300 % and nanoindentation confirms the cohesion mechanism to improve bulk stiffness by around 200 %.
ABSTRACT
A previously introduced tyrosinase-activated polymerization of Tyr- and Cys-bearing peptides yielding artificial mussel-glue proteins is realized without the need of the specific enzyme by a chemical activation route. This decouples the sequence of polymerizable peptides (unimers) from the constraints of tyrosinase substrates and enables the polymerization of minimal motifs such as Dopa-Lys-Cys (Umini *KC ) or Dopa-Gly-Cys (Umini *GC ). In the polymerization procedure, sodium periodate is used to oxidize Dopa residues of the unimers to Dopa-quinones to which the thiol of a Cys residue is added in a Michael-type reaction. The resulting polyUmini *KC and polyUmini *GC exhibit a thiol-catechol connectivity as a potent adhesive functionality at each repeat unit. QCM-D experiments show the excellent substrate adsorption properties of the products from the chemically activated polymerization. On aluminum oxide surfaces, polyUmini *KC rapidly forms a coating, even under seawater model conditions and the coating resists rinsing with hypersaline solution of 4.2 M salt mixtures. While the sodium periodate oxidation is less specific than the tyrosinase reaction and requires the implementation of Dopa instead of Tyr residues into the polymerizable unimers, the chemical route makes scale-up more easily accessible.
Subject(s)
Adhesives/chemistry , Peptides/chemistry , Adhesives/chemical synthesis , Amino Acid Sequence , Dihydroxyphenylalanine/chemistry , Kinetics , Monophenol Monooxygenase/metabolism , Peptides/metabolism , Periodic Acid/chemistry , Polymerization , Proteins/chemistry , Quartz Crystal Microbalance Techniques , Quinones/chemistry , Sulfhydryl Compounds/chemistryABSTRACT
Peptides with suitable substrate properties for a specific tyrosinase are selected by combinatorial means from a one-bead-one-compound (OBOC) peptide library. The identified sequences exhibit tyrosine residues that are rapidly oxidized to 3,4-dihydroxyphenylalanine (Dopa), making the peptides interesting for enzyme-activated adhesives. The selection process of peptides involves tyrosinase oxidation of tyrosine-bearing sequences on a solid support, yielding dopaquinone residues (fish from the sequence pool), to which thiol-functional fluorescent probes attach by Michael-reaction (clip to mark). Labeled supports are isolated and sequence readout is feasible by MALDI-TOF-MS/MS to reveal peptides, while activation kinetics as well as enzyme-activated coating behavior are verifying the proper selection.
ABSTRACT
A novel strategy to generate adhesive protein analogues by enzyme-induced polymerization of peptides is reported. Peptide polymerization relies on tyrosinase oxidation of tyrosine residues to Dopaquinones, which rapidly form cysteinyldopa-moieties with free thiols from cysteine residues, thereby linking unimers and generating adhesive polymers. The resulting artificial protein analogues show strong adsorption to different surfaces, even resisting hypersaline conditions. Remarkable adhesion energies of up to 10.9â mJ m-2 are found in single adhesion events and average values are superior to those reported for mussel foot proteins that constitute the gluing interfaces.
