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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geprüft am 02.05.2024