Nanoscale organization of mitochondrial microcompartments revealed by combining tracking and localization microscopy.
While detailed information on the nanoscale-organization of proteins within intracellular membranes has emerged from electron microcopy, information on their spatiotemporal dynamics is scarce. By use of a photostable rhodamine attached specifically to Halo-tagged proteins in mitochondrial membranes, we were able to track and localize single protein complexes such as Tom20 and ATP synthase in suborganellar structures in live cells. Individual membrane proteins in the inner and outer membrane of mitochondria were imaged over long time periods with localization precisions below 15 nm. Projection of single molecule trajectories revealed diffusion-restricting microcompartments such as individual cristae in mitochondria. At the same time, protein-specific diffusion characteristics within different mitochondrial membranes could be extracted.
Division of Mitochondrial Dynamics, Department of Biology, University of Osnabrück, Barbarastrasse 11, 49076 Osnabrück, Germany.
Timo
T
Appelhans
Christian P
CP
Richter
Verena
V
Wilkens
Samuel T
ST
Hess
Jacob
J
Piehler
Karin B
KB
Busch
D000251:Adenosine Triphosphatases / Q000737:chemistry* / Q000378:metabolism; D002462:Cell Membrane / Q000737:chemistry; D006367:HeLa Cells; D006801:Humans; D026901:Membrane Transport Proteins / Q000737:chemistry*; D008854:Microscopy, Electron; D008928:Mitochondria / Q000737:chemistry*; D000090622:Mitochondrial Precursor Protein Import Complex Proteins; D049329:Nanostructures / Q000737:chemistry*; D011956:Receptors, Cell Surface / Q000737:chemistry*; D012235:Rhodamines / Q000737:chemistry
D026901:Membrane Transport Proteins; D000090622:Mitochondrial Precursor Protein Import Complex Proteins; D011956:Receptors, Cell Surface; D012235:Rhodamines; C084008:TOMM20 protein, human; D000251:Adenosine Triphosphatases