Nanoscopic anatomy of dynamic multi-protein complexes at membranes resolved by graphene-induced energy transfer

Autor(en): Fuellbrunn, Nadia
Li, Zehao
Jorde, Lara
Richter, Christian P.
Kurre, Rainer 
Langemeyer, Lars
Yu, Changyuan
Meyer, Carola 
Enderlein, Jorg 
Ungermann, Christian 
Piehler, Jacob 
You, Changjiang 
Stichwörter: BINDING; Biology; DNA; FUSION; HOPS; IDENTIFICATION; Life Sciences & Biomedicine - Other Topics; LOCALIZATION; MICROSCOPY; ORIENTATION; RAB GTPASE; RECEPTOR INTERACTIONS
Erscheinungsdatum: 2021
Herausgeber: ELIFE SCIENCES PUBLICATIONS LTD
Journal: ELIFE
Volumen: 10
Zusammenfassung: 
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 subnanometer resolution.
ISSN: 2050084X
DOI: 10.7554/eLife.62501

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