Synthesis of Multi-Protein Complexes through Charge-Directed Sequential Activation of Tyrosine Residues.

Site-selective protein-protein coupling has long been a goal of chemical biology research. In recent years, that goal has been realized to varying degrees through a number of techniques, including the use of tyrosinase-based coupling strategies. Early publications utilizing tyrosinase from (abTYR) showed the potential to convert tyrosine residues into -quinone functional groups, but this enzyme is challenging to produce recombinantly and suffers from some limitations in substrate scope.

Tyrosinase-Mediated Oxidative Coupling of Tyrosine Tags on Peptides and Proteins.

Oxidative coupling (OC) through -quinone intermediates has been established as an efficient and site-selective way to modify protein N-termini and the unnatural amino acid -aminophenylalanine (aF). Recently, we reported that the tyrosinase-mediated oxidation of phenol-tagged cargo molecules is a particularly convenient method of generating -quinones in situ. The coupling partners can be easily prepared and stored, the reaction takes place under mild conditions (phosphate buffer, pH 6.