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. 2021 Dec 31;10:e68213. doi: 10.7554/eLife.68213

Figure 1. TMEM223 and SMIM4 interact with the mitochondrial ribosome.

Figure 1.

(A) Mitochondria isolated from cells expressing mL62FLAG were subjected to co-immunoprecipitation. Natively isolated complexes were separated by sucrose density gradient ultracentrifugation. Fractions (1–15) were analyzed by western blotting, using indicated antibodies against components of the 39S mtLSU (mL62, uL1m, uL3m, and uL23m) and 28S mtSSU (uS14m and bS16m) subunits. (B) Mitochondria were isolated from wild-type (WT) and mL62FLAG-expressing cells, cultured in SILAC-medium, and subjected to co-immunoisolation. Eluates were analyzed by quantitative mass spectrometry (LC-MS/MS) (n=4). Ribosomal proteins of the mtLSU and mtSSU are indicated in red and blue, respectively. Dashed lines indicate a p-value of 0.05 and mean mL62FLAG/WT ratios. (C) Scheme of proteins identified in (B). (D) Complexes containing mL62FLAG were purified as in (A) and (B) and analyzed by western blotting (Total, 0.75%; Eluate, 100%).

Figure 1—source data 1. Data of Figure 1A and D.