Nanoscopic anatomy of dynamic multi-protein complexes at membranes resolved by graphene-induced energy transfer
Nadia Füllbrunn,
Zehao Li,
Lara Jorde,
Christian P Richter,
Rainer Kurre,
Lars Langemeyer,
Changyuan Yu,
Carola Meyer,
Jörg Enderlein,
Christian Ungermann,
Jacob Piehler,
Changjiang You
Affiliations
Nadia Füllbrunn
Department of Biology/Chemistry, University of Osnabrück, Osnabrück, Germany; Center of Cellular Nanoanalytics (CellNanOs), University of Osnabrück, Osnabrück, Germany
Zehao Li
Department of Biology/Chemistry, University of Osnabrück, Osnabrück, Germany; Center of Cellular Nanoanalytics (CellNanOs), University of Osnabrück, Osnabrück, Germany; College of Life Sciences, Beijing University of Chemical Technology, Beijing, China
Lara Jorde
Department of Physics, University of Osnabrück, Osnabrück, Germany
Christian P Richter
Department of Biology/Chemistry, University of Osnabrück, Osnabrück, Germany
Rainer Kurre
Department of Biology/Chemistry, University of Osnabrück, Osnabrück, Germany; Center of Cellular Nanoanalytics (CellNanOs), University of Osnabrück, Osnabrück, Germany
Center of Cellular Nanoanalytics (CellNanOs), University of Osnabrück, Osnabrück, Germany; Department of Physics, University of Osnabrück, Osnabrück, Germany
3rd Institute of Physics - Biophysics, Georg August University, Göttingen, Germany; Cluster of Excellence “Multiscale Bioimaging: from Molecular Machines to Networks of Excitable Cells” (MBExC), Georg August University, Göttingen, Germany
Department of Biology/Chemistry, University of Osnabrück, Osnabrück, Germany; Center of Cellular Nanoanalytics (CellNanOs), University of Osnabrück, Osnabrück, Germany
Department of Biology/Chemistry, University of Osnabrück, Osnabrück, Germany; Center of Cellular Nanoanalytics (CellNanOs), University of Osnabrück, Osnabrück, Germany
Department of Biology/Chemistry, University of Osnabrück, Osnabrück, Germany; Center of Cellular Nanoanalytics (CellNanOs), University of Osnabrück, Osnabrück, Germany
Insights into the conformational organization and dynamics of proteins complexes at membranes is essential for our mechanistic understanding of numerous key biological processes. Here, we introduce graphene-induced energy transfer (GIET) to probe axial orientation of arrested macromolecules at lipid monolayers. Based on a calibrated distance-dependent efficiency within a dynamic range of 25 nm, we analyzed the conformational organization of proteins and complexes involved in tethering and fusion at the lysosome-like yeast vacuole. We observed that the membrane-anchored Rab7-like GTPase Ypt7 shows conformational reorganization upon interactions with effector proteins. Ensemble and time-resolved single-molecule GIET experiments revealed that the HOPS tethering complex, when recruited via Ypt7 to membranes, is dynamically alternating between a ‘closed’ and an ‘open’ conformation, with the latter possibly interacting with incoming vesicles. Our work highlights GIET as a unique spectroscopic ruler to reveal the axial orientation and dynamics of macromolecular complexes at biological membranes with sub-nanometer resolution.
