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

Mahdavi, Alborz and Hamblin, Graham D. and Jindal, Granton A. and Bagert, John D. and Dong, Cathy and Sweredoski, Michael J. and Hess, Sonja and Schuman, Erin M. and Tirrell, David A. (2016) Engineered Aminoacyl-tRNA Synthetase for Cell-Selective Analysis of Mammalian Protein Synthesis. Journal of the American Chemical Society, 138 (13). pp. 4278-4281. ISSN 0002-7863. PMCID PMC4825725. http://resolver.caltech.edu/CaltechAUTHORS:20160404-075529150

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Abstract

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.


Item Type:Article
Related URLs:
URLURL TypeDescription
http://dx.doi.org/10.1021/jacs.5b08980DOIArticle
http://pubs.acs.org/doi/abs/10.1021/jacs.5b08980PublisherArticle
http://pubs.acs.org/doi/suppl/10.1021/jacs.5b08980PublisherSupporting Information
https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4825725/PubMed CentralArticle
ORCID:
AuthorORCID
Mahdavi, Alborz0000-0002-8790-8112
Sweredoski, Michael J.0000-0003-0878-3831
Hess, Sonja0000-0002-5904-9816
Tirrell, David A.0000-0003-3175-4596
Additional Information:© 2016 American Chemical Society. This is an open access article published under an ACS AuthorChoice License, which permits copying and redistribution of the article or any adaptations for non-commercial purposes. Received: August 24, 2015. Publication Date (Web): March 18, 2016. We are grateful for financial support by National Institutes of Health grant NIH R01 GM062523 and the Programmable Molecular Technology Initiative of the Gordon and Betty Moore Foundation, the Institute for Collaborative Biotechnologies through grant W911NF-09-0001 from U.S. Army Research Office. M.S. and S.H. were supported by the Gordon and Betty Moore Foundation through grant GMBF775 and NIH grant 1S10RR029594-01A1. A.M. was supported by a scholarship from the National Science and Engineering Research Council of Canada and by a postgraduate scholarship from the Donna and Benjamin M. Rosen Center for Bioengineering at Caltech. G.D.H. was supported by a postdoctoral fellowship from the National Science and Engineering Research Council of Canada. The authors declare no competing financial interest.
Funders:
Funding AgencyGrant Number
NIHR01 GM062523
Gordon and Betty Moore FoundationUNSPECIFIED
Army Research Office (ARO)W911NF-09-0001
Gordon and Betty Moore FoundationGMBF775
NIH1S10RR029594-01A1
Natural Sciences and Engineering Research Council of Canada (NSERC)UNSPECIFIED
Donna and Benjamin M. Rosen Center for BioengineeringUNSPECIFIED
PubMed Central ID:PMC4825725
Record Number:CaltechAUTHORS:20160404-075529150
Persistent URL:http://resolver.caltech.edu/CaltechAUTHORS:20160404-075529150
Official Citation:Engineered Aminoacyl-tRNA Synthetase for Cell-Selective Analysis of Mammalian Protein Synthesis Alborz Mahdavi, Graham D. Hamblin, Granton A. Jindal, John D. Bagert, Cathy Dong, Michael J. Sweredoski, Sonja Hess, Erin M. Schuman, and David A. Tirrell Journal of the American Chemical Society 2016 138 (13), 4278-4281 DOI: 10.1021/jacs.5b08980
Usage Policy:No commercial reproduction, distribution, display or performance rights in this work are provided.
ID Code:65873
Collection:CaltechAUTHORS
Deposited By: Ruth Sustaita
Deposited On:04 Apr 2016 19:45
Last Modified:18 Jul 2017 19:46

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