TY - JOUR

T1 - Shuttling of PINK1 between Mitochondrial Microcompartments Resolved by Triple-Color Superresolution Microscopy

AU - Beinlich, Felix R.M.

AU - Drees, Christoph

AU - Piehler, Jacob

AU - Busch, Karin B

PY - 2015

Y1 - 2015

N2 - 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.

AB - 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.

KW - HeLa Cells

KW - Humans

KW - Membrane Potential, Mitochondrial

KW - Microscopy, Fluorescence

KW - Mitochondria/metabolism

KW - Mitochondrial Membranes/metabolism

KW - Protein Kinases/analysis

KW - Protein Structure, Tertiary

U2 - 10.1021/acschembio.5b00295

DO - 10.1021/acschembio.5b00295

M3 - Letter

C2 - 26046594

VL - 10

SP - 1970

EP - 1976

JO - A C S Chemical Biology

JF - A C S Chemical Biology

SN - 1554-8929

IS - 9

ER -