Engineered Aminoacyl-tRNA Synthetase for Cell-Selective Analysis of Mammalian Protein Synthesis

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• 作者：Alborz Mahdavi ; Graham D. Hamblin ; Granton A. Jindal ; John D. Bagert ; Cathy Dong ; Michael J. Sweredoski ; Sonja Hess ; Erin M. Schuman ; David A. Tirrell
• 刊名：Journal of the American Chemical Society
• 出版年：2016
• 出版时间：April 6, 2016
• 年：2016
• 卷：138
• 期：13
• 页码：4278-4281
• 全文大小：311K
• ISSN：1520-5126

文摘

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.