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Cell-type-specific metabolic labeling of nascent proteomes in vivo

Alvarez-Castelao, Beatriz and Schanzenbächer, Christoph T. and Hanus, Cyril and Glock, Caspar and tom Dieck, Susanne and Dörrbaum, Aline R. and Bartnik, Ina and Nassim-Assir, Belquis and Ciirdaeva, Elena and Mueller, Anke and Dieterich, Daniela C. and Tirrell, David A. and Langer, Julian D. and Schuman, Erin M. (2017) Cell-type-specific metabolic labeling of nascent proteomes in vivo. Nature Biotechnology, 35 (12). pp. 1196-1201. ISSN 1087-0156. https://resolver.caltech.edu/CaltechAUTHORS:20171113-085844680

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Abstract

Although advances in protein labeling methods have made it possible to measure the proteome of mixed cell populations, it has not been possible to isolate cell-type-specific proteomes in vivo. This is because the existing methods for metabolic protein labeling in vivo access all cell types. We report the development of a transgenic mouse line where Cre-recombinase-induced expression of a mutant methionyl-tRNA synthetase (L274G) enables the cell-type-specific labeling of nascent proteins with a non-canonical amino-acid and click chemistry. Using immunoblotting, imaging and mass spectrometry, we use our transgenic mouse to label and analyze proteins in excitatory principal neurons and Purkinje neurons in vitro (brain slices) and in vivo. We discover more than 200 proteins that are differentially regulated in hippocampal excitatory neurons by exposing mice to an environment with enriched sensory cues. Our approach can be used to isolate, analyze and quantitate cell-type-specific proteomes and their dynamics in healthy and diseased tissues.


Item Type:Article
Related URLs:
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https://doi.org/10.1038/nbt.4016DOIArticle
https://www.nature.com/articles/nbt.4016PublisherArticle
http://rdcu.be/yrqsPublisherFree ReadCube access
ORCID:
AuthorORCID
Tirrell, David A.0000-0003-3175-4596
Additional Information:© 2017 Macmillan Publishers Limited, part of Springer Nature. Received: 12 December 2016; Accepted: 19 October 2017; Published online: 06 November 2017. We thank H. Geptin, D. Vogel, N. Fuerst, I. Wüllenweber and F. Rupprecht for excellent technical assistance. We thank E. Noll for the synthesis of ANL and P. Landgraf for the synthesis of the SH-alkyne. We thank S. Garg for her help with FUNCAT and M. Heumueller for his help with some of the experiments. We thank R. Pieaud and S. Junek for their assistance with imaging. We thank F. Kretschmer for his help with the open field analysis. We thank E. Northrup, S. Zeissler, S. Gil Mast and the animal facility of MPI for Brain Research for their excellent support. We thank J. Sanes and J. Chakkalakal for the early generation of a Thy-1 MetRS* mouse. Work in the laboratory of E.M.S. is supported by the Max Planck Society, the European Research Council, DFG CRC 902 and 1080, and the DFG Cluster of Excellence for Macromolecular Complexes. B.A. was supported by a Marie Curie Intra-European Fellowship for career development. C.H. was supported by a Marie Curie Career Integration Grant. D.C.D. is supported by DFG CRC 779 and 854. Research on proteomic labelling at Caltech is supported by the Programmable Molecular Technology Initiative of the Gordon and Betty Moore Foundation. Author Contributions: B.A.-C., C.T.S. and C.H.: conception and design of experiments, acquisition, analysis and interpretation of data. C.G., S.t.D. and A.R.D.: conception and design of experiments, acquisition of data. I.B., B.N.-A. and E.C.: acquisition of data. A.M.: provided reagents. D.D.: provided reagents and advice on experiments. D.A.T.: conception and design of experiments. J.D.L.: acquisition of data, analysis and interpretation of data. E.M.S.: conception and design of experiments, analysis and interpretation of data, drafting, writing and revising the article. All authors contributed to the writing and revision of the manuscript. The authors declare no competing financial interests.
Funders:
Funding AgencyGrant Number
Max Planck SocietyUNSPECIFIED
European Research Council (ERC)UNSPECIFIED
Deutsche Forschungsgemeinschaft (DFG)CRC 902
Deutsche Forschungsgemeinschaft (DFG)CRC 1080
Marie Curie FellowshipUNSPECIFIED
Deutsche Forschungsgemeinschaft (DFG)CRC 779
Deutsche Forschungsgemeinschaft (DFG)CRC 854
Gordon and Betty Moore FoundationUNSPECIFIED
Issue or Number:12
Record Number:CaltechAUTHORS:20171113-085844680
Persistent URL:https://resolver.caltech.edu/CaltechAUTHORS:20171113-085844680
Usage Policy:No commercial reproduction, distribution, display or performance rights in this work are provided.
ID Code:83141
Collection:CaltechAUTHORS
Deposited By: Tony Diaz
Deposited On:14 Nov 2017 23:25
Last Modified:03 Oct 2019 19:02

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