Expanding the natural repertoire of protein structure and function

This review considers chemical and genetic approaches to the modification of protein structure. The historical interest in chemical and site-directed modifications will be briefly covered. Current chemical modification strategies will be presented. Biosynthetic mutagenesis with unnatural aminoacyl-tRNAs and current synthetic peptide ligation technologies will be covered in greater detail. The application of combinatorial genetic methods (e.g. phage display, DNA shuffling) to protein engineering with unnatural amino acids will be briefly discussed, with emphasis on the in vitro evolution of new enzymatic function (i.e. aminoacyl-tRNA synthetases). Throughout the review, the powerful insights gained from the combined use of these technologies will be illustrated by examples that focus on the elucidation of protein-ligand interactions.