Our group aims to understand and monitor the dynamic formation of protein complexes in the context of membrane dynamics. For example, respiratory complexes of mitochondria that are arranged in a sequence of electron transfer reactions form rather stable supercomplexes (Rieger et al., 2017) in response to specific (energetic) states. On the other hand, membrane proteins transiently interact with soluble proteins, e.g. during membrane fission and fusion such as exocytosis. The interaction of membrane - membrane protein complexes or membrane – soluble protein complexes often is spatially and temporally restricted and thus difficult to survey. Yet, specific and spatially confined nano-organization mutually is prerequisite for local functionality (Beinlich et al., 2015). In our group, we use and implement diverse fluorescence-based microscopy techniques ranging from fluorescence lifetime imaging microscopy (FLIM) (Söhnel et al., 2016; Rieger et al., 2017) to superresolution imaging (Appelhans and Busch, 2017, 2017; Appelhans et al., 2012; Beinlich et al., 2015; Wilmes et al.) in order to determine and analyze the formation and localization of membrane protein complexes in their native environment linked to processes associated with cellular adaptation or differentiation.
- Appelhans, T., and Busch, K. (2017a). Single Molecule Tracking and Localization of Mitochondrial Protein Complexes in Live Cells. Methods Mol. Biol. 1567, 273–291.
- Appelhans, T., and Busch, K.B. (2017b). Dynamic imaging of mitochondrial membrane proteins in specific sub-organelle membrane locations. Biophysical Reviews 9, 345–352.
- Appelhans, T., Richter, C., Wilkens, V., Hess, S., Piehler, J., and Busch, K. (2012). Nanoscale organization of mitochondrial microcompartments revealed by combining tracking and localization microscopy. Nano Letters 12, 610–616.
- Beinlich, F., Drees, C., Piehler, J., and Busch, K. (2015). Shuttling of PINK1 between Mitochondrial Microcompartments Resolved by Triple-Color Superresolution Microscopy. ACS Chemical Biology 10, 1970–1976.
- Rieger, B., Shalaeva, D.N., Söhnel, A.-C.C., Kohl, W., Duwe, P., Mulkidjanian, A.Y., and Busch, K.B. (2017). Lifetime imaging of GFP at CoxVIIIa reports respiratory supercomplex assembly in live cells. Sci Rep 7, 46055.
- Söhnel, A.-C.C., Kohl, W., Gregor, I., Enderlein, J., Rieger, B., and Busch, K.B. (2016). Probing of protein localization and shuttling in mitochondrial microcompartments by FLIM with sub-diffraction resolution. Biochim. Biophys. Acta 1857, 1290–1299.
- Wilmes, S, Staufenbiel, M, Lisse, D, Richter, CP, Beutel, O, Busch, KB, Hess, ST, and Piehler, J Triple-color super-resolution imaging of live cells: resolving submicroscopic receptor organization in the plasma membrane. Angewandte Chemie 51, 4868–4871.