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

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.

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Item Permalink: https://hdl.handle.net/21.11116/0000-000B-216A-9 Version Permalink: https://hdl.handle.net/21.11116/0000-000B-216B-8
Genre: Journal Article

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https://pubmed.ncbi.nlm.nih.gov/35499346/ (Any fulltext)
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 Creators:
Nassim-Assir, Belquis1, Author
Corredera, Daniel Ouro2, Author
Alvarez-Pardo, Rodrigo2, Author
Dieck, Susanne Tom1, Author
Ciirdaeva, Elena1, Author
Schuman, Erin M1, Author
Alvarez-Castelao, Beatriz2, Author
Affiliations:
1Max Planck Institute for Brain Research, Max Planck Society, Max-von-Laue-Str. 4, 60438 Frankfurt am Main, DE, ou_2461692              
2 Department of Biochemistry and Molecular Biology, Veterinary School, Complutense University of Madrid., ou_persistent22              

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 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.

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Language(s): eng - English
 Dates: 2022-04-22
 Publication Status: Published online
 Pages: -
 Publishing info: -
 Table of Contents: -
 Rev. Type: -
 Identifiers: DOI: 10.3791/63713
PMID: 35499346
 Degree: -

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Title: J. Vis. Exp.
  Other : Journal of Visualized Experiments: JoVE
  Abbreviation : J. Vis. Exp.
Source Genre: Journal
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Publ. Info: Rockville Pike, Bethesda MD : JoVE
Pages: - Volume / Issue: 182 Sequence Number: - Start / End Page: - Identifier: ISSN: 1940-087X
CoNE: https://pure.mpg.de/cone/journals/resource/1940087X