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. 2012 May 28;197(5):595–604. doi: 10.1083/jcb.201110047

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© 2012 Gebert et al.

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Figure 1. — Mgr2 is a subunit of the TIM23 complex. (A) Yeast cells were labeled with [¹³C₆]lysine/[¹³C₆]arginine (wild type) or their ¹²C-containing counterparts (Tim21_ProtA strain). Mitochondria were isolated, mixed, and lysed with digitonin. The TIM23 complex was isolated by IgG affinity purification. The mean log₁₀ L/H ratio was plotted against the corresponding p-value (−log₁₀) of each protein. Red, presequence translocase subunits including Mgr2; green, complexes III and IV; yellow, mitochondrial carrier family; brown, Tim44; black and grey, further proteins. See Materials and methods and Table S1. (B) Wild-type (WT) and Tim23_ProtA mitochondria were lysed in digitonin and incubated with IgG Sepharose. Bound complexes were eluted by TEV protease cleavage and analyzed by SDS-PAGE and immunoblotting. Eluate: 100%; load: TIM/PAM subunits 17%, respiratory chain subunits 0.5%, and further proteins 6%. The arrowhead indicates the signal of Tim23_ProtA. (C) Wild-type and Tim21_ProtA mitochondria were analyzed as described for B. Eluate: 100%; load: TIM/PAM subunits 6%, respiratory chain subunits 0.5%, and further proteins 4%. The arrowhead indicates the signal of Tim21_ProtA.