Impact of Nonnatural Amino Acid Mutagenesis on the in Vivo Function and Binding Modes of a Transcriptional Activator

摘要

Given the central role of protein-protein interactions in biological functions, the development of methods for the discovery and characterization of such interactions has been of paramount importance in the scientific community. Ideally these methods are compatible with live-cell studies so that protein-protein interactions can be investigated in their native context. Toward that end, recent advances in nonsense suppression technology have enabled the site-specific incorporation of amino acids bearing moieties for photo-activatable cross-linking reactions into proteins in bacteria, yeast, and human cells. However, there have been few applications of this technology in eukaryotes, perhaps due to challenges in obtaining efficient incorporation of the nonnatural amino acids as well as limited knowledge about the impact of these changes on structurernand function.