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Each mitochondrial compartment contains varying protein compositions that underlie a diversity of localized functions. Insights into the localization of mitochondrial intermembrane space bridging (MIB) components will have an impact on our understanding of mitochondrial architecture, dynamics and function. Using the novel visualizable genetic tags miniSOG and APEX2 in cultured mouse cardiac and human astrocyte cell lines and electron tomography, we have mapped at nanoscale resolution three key MIB components: Mic19, Mic60 and Sam50, in the environment of structural landmarks such as cristae and crista junctions (CJs). Tagged Mic19 and Mic60 were located at CJs, distributed in a network pattern along the mitochondrial periphery, and also enriched inside cristae. We discovered an association of Mic19 with cytochrome c oxidase subunit IV. It was also found that tagged Sam50 is not uniformly distributed in the outer mitochondrial membrane and appears to incompletely overlap with Mic19 or Mic60-positive domains, most notably at the CJs.

Sub-mitochondrial localization of genetic-tagged MIB interacting partners: Mic19, Mic60 and Sam50