In JoVE (1)
Other Publications (1)
Articles by Felix R.M. Beinlich in JoVE
Multi-color Localization Microscopy of Single Membrane Proteins in Organelles of Live Mammalian Cells Timo Appelhans1, Felix R.M. Beinlich1, Christian P. Richter1, Rainer Kurre2, Karin B. Busch1,3 1School of Biology, University of Osnabrück, 2Center of Cellular Nanoanalytics, Integrated Bioimaging Facility, University of Osnabrück, 3Department of Biology, WWU Münster Here, we present a protocol for multi-color localization of single membrane proteins in organelles of live cells. To attach fluorophores, self-labeling proteins are used. Proteins, located in different membranes compartments of the same organelle, can be localized with a precision of ~18 nm.
Other articles by Felix R.M. Beinlich on PubMed
Shuttling of PINK1 Between Mitochondrial Microcompartments Resolved by Triple-Color Superresolution Microscopy ACS Chemical Biology. Sep, 2015 | Pubmed ID: 26046594 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.