Engineered Aminoacyl-tRNA Synthetase for Cell-Selective Analysis of Mammalian Protein Synthesis
| Autor: | John D. Bagert, Alborz Mahdavi, Erin M. Schuman, Graham D. Hamblin, Cathy Dong, Michael J. Sweredoski, Sonja Hess, Granton A. Jindal, David A. Tirrell |
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| Rok vydání: | 2016 |
| Předmět: |
0301 basic medicine
RNA Transfer Met Proteome Cell Mutant Nanotechnology Methionine-tRNA Ligase 010402 general chemistry 01 natural sciences Biochemistry Catalysis Amino Acyl-tRNA Synthetases Mice 03 medical and health sciences chemistry.chemical_compound Colloid and Surface Chemistry Cricetinae Escherichia coli medicine Protein biosynthesis Animals Humans Amino Acids Mammals chemistry.chemical_classification Chemistry Aminoacyl tRNA synthetase Communication Haplorhini General Chemistry 0104 chemical sciences Amino acid 030104 developmental biology medicine.anatomical_structure Cell culture Protein Biosynthesis Transfer RNA HeLa Cells |
| Zdroj: | Journal of the American Chemical Society |
| ISSN: | 1520-5126 0002-7863 |
| DOI: | 10.1021/jacs.5b08980 |
| Popis: | Methods for cell-selective analysis of proteome dynamics will facilitate studies of biological processes in multicellular organisms. Here we describe a mutant murine methionyl-tRNA synthetase (designated L274GMmMetRS) that charges the noncanonical amino acid azidonorleucine (Anl) to elongator tRNA^(Met) 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 does not 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. |
| Databáze: | OpenAIRE |
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