Cited by 220Advanced Proteomics Techniques and ApplicationsClick Chemistry and ApplicationsBiotin and Related Studies

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

Alvarez-Castelao, Beatriz, Schanzenbächer, Christoph T, Hanus, Cyril, Glock, Caspar, tom Dieck, Susanne, Dörrbaum, Aline R, Bartnik, Ina, Nassim-Assir, Belquis +6 more·2017·Nature Biotechnology

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.

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Crossref · 10.1038/nbt.4016

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Authors
Alvarez-Castelao, B., Schanzenbächer, C. T., Hanus, C., Glock, C., tom Dieck, S., Dörrbaum, A. R., Bartnik, I., Nassim-Assir, B., Ciirdaeva, E., Mueller, A., Dieterich, D. C., Tirrell, D. A., Langer, J. D., & Schuman, E. M.
Affiliations
  • Max Planck Institute of Biophysics (DE)
  • Max Planck Institute for Brain Research (DE)
  • Center for Behavioral Brain Sciences (DE)
  • California Institute of Technology (US)
Corresponding author
Schuman, Erin M
Venue
Nature Biotechnology
Published
2017-11-06
Volume
35
Issue
12
Pages
1196-1201
ISSN
  • 0733-222X (ISSN-L)
  • 1087-0156
  • 1546-1696
OpenAlex
W2765371514
Cited by
219
References
50
Citations by year
2026: 15 · 2025: 30 · 2024: 40 · 2023: 27 · 2022: 25 · 2021: 19 · 2020: 26 · 2019: 17 · 2018: 17 · 2017: 2
FWCI
8.38
Type
journal article
Language
en
Open access
closed
Locations
  • Nature Biotechnology (publishedVersion)
  • PubMed (publishedVersion)
  • CaltechAUTHORS (California Institute of Technology) (submittedVersion)
  • Max Planck Digital Library (submittedVersion)
  • MPG.PuRe (Max Planck Society) (submittedVersion)
Funders
Agence Nationale de la Recherche (ANR-16-CE16-0009)
Topics
  • Physical Sciences › Chemistry › Spectroscopy › Advanced Proteomics Techniques and Applications
  • Physical Sciences › Chemistry › Organic Chemistry › Click Chemistry and Applications
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Cell Biology › Biotin and Related Studies
Keywords
click chemistry, transgenic mice, Cre recombinase, Purkinje cells, hippocampus
MeSH terms
Amino Acids, Animals, Female, Gene Expression Regulation, Male, Methionine-tRNA Ligase, Mice, Transgenic, Neurons, Integrases, Proteome, Proteomics, Mice, Click Chemistry

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