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

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

Journal of visualized experiments : JoVE 2022 Apr 13

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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.

Citation

Belquis Nassim-Assir, Daniel Ouro Corredera, Rodrigo Alvarez-Pardo, Susanne Tom Dieck, Elena Ciirdaeva, Erin M Schuman, Beatriz Alvarez-Castelao. Cell-type Specific Protein Purification and Identification from Complex Tissues using a Mutant Methionine tRNA Synthetase Mouse Line. Journal of visualized experiments : JoVE. 2022 Apr 13(182)