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 - 2015/9/18
Y1 - 2015/9/18
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
SN - 1554-8929
VL - 10
SP - 1970
EP - 1976
JO - A C S Chemical Biology
JF - A C S Chemical Biology
IS - 9
ER -