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Single-Molecule Metal-Induced Energy Transfer (smMIET)

Project description

Probing protein structure and dynamics at the nanoscale

First presented in 2012, single-molecule metal-induced energy transfer can be used to determine the position of individual molecules near a metal surface with nanometre accuracy. When a fluorescent molecule is brought close to a metal surface, it transfers its excited state energy to surface plasmons in the metal. Researchers have been using this method for mapping cellular membranes with nanometre axial resolution. The aim of the EU-funded smMIET project is to expand the potential applications of this novel single-molecule localisation concept. In particular, the project will apply the technology for resolving the global structure of macromolecular complexes, the conformational fluctuations of intrinsically disordered proteins, the dynamics of lipid membranes, or the transport of proteins across lipid bilayers.

Objective

The core aim of the project is to develop the technology of Single-Molecule Metal-Induced Energy Transfer (smMIET) for resolving macromolecular structure and dynamics with sub-nanometre spatial resolution and nanosecond temporal resolution. Metal-Induced Energy Transfer or MIET was first developed in our group in 2012 for mapping cellular membranes with nanometre axial resolution. It exploits the effect that a fluorescent molecule, when brought close to a metal surface, can transfer its excited state energy to surface plasmons in the metal, which leads to a strong distance-dependence of its fluorescence lifetime and intensity. This strong lifetime-distance dependence allows for converting a measured fluorescence lifetime into a distance from the metal surface. Combining this concept with single-molecule localization super-resolution microscopy and with fluorescence correlation spectroscopy will resolve three-dimensional structures with nanometre isotropic resolution, and structural dynamics on the nanometre length scale with nanosecond temporal resolution. Among its many applications, the project will develop and apply smMIET for resolving the global structure of macromolecular complexes and its dynamics, the conformational fluctuations of intrinsically disordered proteins, the dynamics of lipid membranes in a leaflet-resolved manner, or the transport of proteins across lipid bilayers. We will establish smMIET as a toolbox for structural and molecular biology that is comparable and complementary in its usefulness and versatility to conventional Förster Resonance Energy Transfer (FRET) or Fluorescence Correlation Spectroscopy (FCS).

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Coordinator

GEORG-AUGUST-UNIVERSITAT GOTTINGEN STIFTUNG OFFENTLICHEN RECHTS
Net EU contribution
€ 2 740 188,00
Address
Wilhelmsplatz 1
37073 Gottingen
Germany

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Region
Niedersachsen Braunschweig Göttingen
Activity type
Higher or Secondary Education Establishments
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Other funding
€ 0,00

Beneficiaries (1)