Engineered Aminoacyl-tRNA Synthetase for Cell-Selective Analysis of Mammalian Protein Synthesis
Abstract
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.
Document Details
- Document Type
- Technical Report
- Publication Date
- Mar 18, 2016
- Accession Number
- AD1052506
Entities
People
- Alborz Mahdavi
- Cathy Dong
- David A. Tirrell
- Erin M. Schuman
- Graham D. Hamblin
- Granton A. Jindal
- John D. Bagert
- Michael J. Sweredoski
- Sonja Hess
Organizations
- California Institute of Technology