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

被引：147

作者：

Alvarez-Castelao, Beatriz ^{[1
]}

Schanzenbaecher, Christoph T. ^{[1
,2
]}

Hanus, Cyril ^{[1
,7
]}

Glock, Caspar ^{[1
]}

Dieck, Susanne Tom ^{[1
]}

Doerrbaum, Aline R. ^{[1
,2
]}

Bartnik, Ina ^{[1
]}

Nassim-Assir, Belquis ^{[1
]}

Ciirdaeva, Elena ^{[1
]}

Mueller, Anke ^{[3
,4
,5
]}

Dieterich, Daniela C. ^{[3
,4
,5
]}

Tirrell, David A. ^{[6
]}

Langer, Julian D. ^{[1
,2
]}

Schuman, Erin M. ^{[1
]}

机构：

[1] Max Planck Inst Brain Res, Frankfurt, Germany

[2] Max Planck Inst Biophys, Frankfurt, Germany

[3] Otto von Guericke Univ, Inst Pharmacol & Toxicol, Magdeburg, Germany

[4] Leibniz Inst Neurobiol, Magdeburg, Germany

[5] Ctr Behav Brain Sci, Magdeburg, Germany

[6] CALTECH, Div Chem & Chem Engn, Pasadena, CA 91125 USA

[7] Inserm 894, Ctr Psychiat & Neurosci, Paris, France

来源：

NATURE BIOTECHNOLOGY | 2017年 / 35卷 / 12期

基金：

欧洲研究理事会;

关键词：

NEWLY SYNTHESIZED PROTEINS; TRANSFER-RNA SYNTHETASE; IDENTIFICATION; EXPRESSION; VISUALIZATION; HIPPOCAMPUS; DISRUPTION; SILAC; LINES;

D O I：

10.1038/nbt.4016

中图分类号：

Q81 [生物工程学（生物技术）]; Q93 [微生物学];

学科分类号：

071005 ; 0836 ; 090102 ; 100705 ;

摘要：

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 Crerecombinase-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.

引用

页码：1196 / +

页数：9

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[1] High-throughput spatial mapping of single-cell RNA-seq data to tissue of origin [J].