In this work we seek to exploit site-specific genetic incorporation of biorthogonal “click” handles or initiating species to generate “grafting-to” and “grafting-from” bioconjugates. Proteins of interest include a robust super-folder green fluorescent protein (sfGFP) and the biologically active, neuronal-targeting protein tetanus toxin fragment C (Hc). Utilizing mutant Methanococcus jannashii synthetase/tRNA pairs specific for non-canonical amino acids (ncAAs) of unique functionality, allows for facile expression of mutant proteins with site-specific handles in E. coli. Azide-bearing ncAA mutant proteins are used to generate unique, dually-functionalized protein-polymer materials via a mild, catalyst-free cycloaddition with bisDBCO-PEG. In contrast to the “grafting-from” method, genetic installation of an ATRP-initiating ncAA allows for growth of block-polymers from the surface of proteins in a similarly site-specific manner. Self-assembled structures can then be made from the mutant protein via polymerization-induced self-assembly (PISA). These techniques, aided by genetic manipulation, produces protein-polymer materials of superior homogeneity. Currently, we are developing protocols for bioconjucation, using both grafting to and grafting from, and ultimate study of mutant sfGFP and Hc-polymer systems.

Investigation of site-specific polymerization techniques via genetic incorporation of bioconjugate handles for studies with self-assembling and stimuli-responsive protein-polymer materials

2019 Spring National Meeting

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