Extended Data Fig. 4: Structures of the TOM complex and its substrate in the supercomplex.
From: Dynamic TOM–TIM23 supercomplex directs mitochondrial protein translocation and sorting
a, Structural comparison of Tom7 in the idle structure (Mandy’s pink, PDB ID: 6UCU) and the active structure (forest green). The two copies of the subunit in a dimeric TOM complex are both shown in a side view and a top view. The phospholipid clamped by Tom7 and Tom40 in the active structure is shown as yellow balls and sticks with transparent density. b, Structural model of an anti-GFP Macrobody. MBP and the anti-GFP nanobody are colored khaki and plum, respectively. c, Purification of the supercomplex complexed with the Macrobody. The elution fractions (grey shaded) from size exclusion chromatography (left) are analyzed by BN-PAGE (top right) and SDS-PAGE (bottom right). BN-PAGE is stained with Coomassie blue. SDS-PAGE is subjected to immunoblotting with an anti-His tag antibody for detection of the Macrobody and Tom22. d, Cryo-EM analysis of the supercomplex with the Macrobody. The raw image (left), 2D classification (middle), and 3D reconstruction (right) are shown. The GFP density in the 2D classes is marked by green arrows. The resolution estimations are from the outputs of 3D classification using RELION. e, Focused 3D classification of the substrate in the Tom40 channel. The six classes are the calculation results from Extended Data Fig. 1e. The three representative classes are also shown in Fig. 1d–f.
