Engineered Aminoacyl-tRNA Synthetase for Cell-Selective Analysis of Mammalian Protein Synthesis
Journal Article - Open Access
California Institute of Technology Pasadena United States
Pagination or Media Count:
Methods for cell-selective analysis of proteome dynamics will facilitate studies of biological processes in multicellular organisms. Here we describe amutant murine methionyl-tRNA synthetase designated L274GMmMetRS that charges the noncanonical amino acid azidonorleucine Anl to elongator tRNAMet in hamster CHO, monkey COS7, and human HeLa cell lines. Proteins made in cells that express the synthetase can be labeled with Anl, tagged with dyes or affinity reagents, and enriched on affinity resin to facilitate identification by mass spectrometry. The method doesnot require expression of orthogonal tRNAs or depletion of canonical amino acids. Successful labeling of proteins with Anl in several mammalian cell lines demonstrates the utility of L274GMmMetRS as a tool for cell-selective analysis of mammalian protein synthesis.