Fig. 4: TOM complex is an exit channel for retrotranslocation of MAD substrates. | Communications Biology

Fig. 4: TOM complex is an exit channel for retrotranslocation of MAD substrates.

From: Tom40 functions as a channel for protein retrotranslocation in the mitochondria-associated degradation (MAD) pathway

Fig. 4: TOM complex is an exit channel for retrotranslocation of MAD substrates.The alt text for this image may have been generated using AI.

A Representative western blots of mitochondrial pellet after releasing (Pellet) and released proteins (Released) from isolated mitochondria from wild-type (WT), doa1Δ, tom5Δ, and doa1Δ tom5Δ cells. The blot was probed with antibodies against GFP for detection of Kgd1p, and antibodies against Tom40p and Cyb2p. Total protein load was assessed using TCE. B–D Quantitation of levels of Kgd1p (B), Tom40p (C) and Cyb2p (D) in mitochondria in (A). E Quantification of mitochondrial proteins released from isolated mitochondria in (A). For panels B–E, signals were first normalized to pellet Cyb2p signals and then normalized to WT. (For panels B–E, n = 4, 1-way ANOVA with Sidak’s multiple comparison test, **p < 0.01; ***p < 0.001; ****p < 0.0001; ns no significance). F, H Representative western blots of mitochondrial pellet after releasing (Pellet) and released proteins (Released) from isolated mitochondria from WT (+) and doa1Δ (−) cells in the presence or absence of 1 mM mPEG. The blot was probed with antibodies against GFP for detection of Kgd1p (F) or Pim1p (H), and against Tom40p and Cyb2p. Total protein load was assessed using TCE. G, I Quantification of released mitochondrial proteins from isolated mitochondria in (F) and (H), respectively (n = 4, 1-way ANOVA with Sidak’s multiple comparison test, *p < 0.05; ns no significance). J, L Representative western blots of mitochondrial pellet after releasing (Pellet) and released proteins (Released) from isolated mitochondria from tom40130/138C (+) and tom40130/138C doa1Δ (−) cells in the presence or absence of 1 mM mPEG. The blot was probed with antibodies against GFP for detection of Kgd1p (J) or Pim1p (L), and against Tom40p and Cyb2p. Total protein load was assessed using TCE. K, M Quantification of released mitochondrial proteins from isolated mitochondria in (J, L) (n = 4, 1-way ANOVA with Sidak’s multiple comparison test, *p < 0.05; **p < 0.01; ***p < 0.001; ns no significance). In all quantification, signals were first normalized to pellet Cyb2p signals and then normalized to WT without mPEG treatment.

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