Fig. 3: ICS proteome mapping using AGK- and TMEM177-APEX2. | Nature Communications

Fig. 3: ICS proteome mapping using AGK- and TMEM177-APEX2.

From: Intracristal space proteome mapping using super-resolution proximity labeling with isotope-coded probes

Fig. 3

a Schematic representation showing TMEM177-APEX2 and AGK-APEX2 labeling using isotope-coded phenol probes for APEX labeling (ICAX) for the identification of intracristal space (ICS) and outer ICS (OCS) proteins, respectively. b Streptavidin-HRP western blot pattern of light desthiobiotin-phenol (LDBP)- and heavy desthiobiotin-phenol (HDBP) proteins labeled with AGK-APEX2 and TMEM177-APEX2. APEX2 expression levels were detected using an anti-V5 antibody. c Heat map-based comparison of the TMEM177/AGK (n = 4 biological replicates) and SCO1/TDRKH datasets. The ICS, OCS, and outer mitochondrial membrane (OMM)/Cytosol groups were classified based on fold-change values; the inner mitochondrial membrane, intermembrane space, and OMM proteins in each group are illustrated according to their known topology in the right panel, with modification sites from the TMEM177/AGK dataset. A blank space indicates that no proteins were detected in the primary SCO1/TDRKH dataset. d Confocal images of TMEM126B-APEX2 and EndoG-APEX2 obtained after immunostaining. Scale bars represent 10 µm. e Localization of TMEM126B and EndoG validated through APEX electron microscopy imaging.

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