Cell-type Specific Protein Purification and Identification from Complex Tissues 
using a Mutant Methionine tRNA Synthetase Mouse Line

Nassim-Assir, Belquis; Corredera, Daniel Ouro; Alvarez-Pardo, Rodrigo; Dieck, Susanne Tom; Ciirdaeva, Elena; Schuman, Erin M; Alvarez-Castelao, Beatriz

doi:10.3791/63713

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Journal Article

Cell-type Specific Protein Purification and Identification from Complex Tissues using a Mutant Methionine tRNA Synthetase Mouse Line

MPS-Authors

Nassim-Assir, Belquis
Max Planck Institute for Brain Research, Max Planck Society;

Dieck, Susanne Tom
Max Planck Institute for Brain Research, Max Planck Society;

Ciirdaeva, Elena
Max Planck Institute for Brain Research, Max Planck Society;

Schuman, Erin M
Max Planck Institute for Brain Research, Max Planck Society;

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https://pubmed.ncbi.nlm.nih.gov/35499346/
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Citation

Nassim-Assir, B., Corredera, D. O., Alvarez-Pardo, R., Dieck, S. T., Ciirdaeva, E., Schuman, E. M., et al. (2022). Cell-type Specific Protein Purification and Identification from Complex Tissues using a Mutant Methionine tRNA Synthetase Mouse Line. J. Vis. Exp., 182. doi:10.3791/63713.

Cite as: https://hdl.handle.net/21.11116/0000-000B-216A-9

Abstract

Understanding protein homeostasis in vivo is key to knowing how the cells work in both physiological and disease conditions. The present protocol describes in vivo labeling and subsequent purification of newly synthesized proteins using an engineered mouse line to direct protein labeling to specific cellular populations. It is an inducible line by Cre recombinase expression of L274G-Methionine tRNA synthetase (MetRS*), enabling azidonorleucine (ANL) incorporation to the proteins, which otherwise will not occur. Using the method described here, it is possible to purify cell-type-specific proteomes labeled in vivo and detect subtle changes in protein content due to sample complexity reduction.