title: Shuttling of PINK1 between Mitochondrial Microcompartments Resolved by Triple-Color Superresolution Microscopy.
abstract: The cytosolic phosphatase and tensin homologue Pten-kinase PINK1 involved in mitochondrial quality control undergoes a proteolytic process inside mitochondria. It has been suggested that the protein is not fully imported into mitochondria during this maturation. Here, we have established live cell triple-color super-resolution microscopy by combining FPALM and tracking and localization microscopy (TALM) in order to unravel the spatiotemporal organization of the C-terminal kinase domain of PINK1 during this process. We find that the kinase domain is imported into active mitochondria and colocalizes with respiratory complex I at the inner mitochondrial membrane. When the processing step inside mitochondria is inhibited or mitochondria are de-energized, full length PINK1 distributes between the outer and the inner mitochondrial membranes, indicating a holdup of import. These findings give the molecular base for a dual role of PINK1-inside energized mitochondria and outside of de-energized mitochondria.
authors.affiliation: Mitochondrial Dynamics Group and Biophysics Group, School of Biology, University of Osnabrück , 49076 Osnabrück, Lower Saxony, Germany.
authors.forename: Felix R M
authors.initials: FR
authors.lastname: Beinlich
authors.affiliation: Mitochondrial Dynamics Group and Biophysics Group, School of Biology, University of Osnabrück , 49076 Osnabrück, Lower Saxony, Germany.
authors.forename: Christoph
authors.initials: C
authors.lastname: Drees
authors.affiliation: Mitochondrial Dynamics Group and Biophysics Group, School of Biology, University of Osnabrück , 49076 Osnabrück, Lower Saxony, Germany.
authors.forename: Jacob
authors.initials: J
authors.lastname: Piehler
authors.affiliation: Mitochondrial Dynamics Group and Biophysics Group, School of Biology, University of Osnabrück , 49076 Osnabrück, Lower Saxony, Germany.
authors.forename: Karin B
authors.initials: KB
authors.lastname: Busch
mesh_terms: D006367:HeLa Cells; D006801:Humans; D053078:Membrane Potential, Mitochondrial; D008856:Microscopy, Fluorescence; D008928:Mitochondria / Q000378:metabolism* / Q000648:ultrastructure*; D051336:Mitochondrial Membranes / Q000378:metabolism / Q000648:ultrastructure; D011494:Protein Kinases / Q000032:analysis* / Q000378:metabolism*; D017434:Protein Structure, Tertiary; D000099202:PTEN-Induced Putative Kinase
chemical_list: D011494:Protein Kinases; D000099202:PTEN-Induced Putative Kinase