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- Project 1: Molecular insights into the role of lipids and regulatory proteins on sodium
- Project 2: NMR based structural and dynamic investigation of the transport cycle of NSS transporters
- Project 3: Purification, functional characterization and reconstitution of DAT and LeuT
- Project 4: Development of a comprehensive model of GABA transporter function
- Project 5: Electrophysiological characterization of neurotransmitter transporters and formulation of an overall kinetic model
- Project 6: Electrophysiological approaches to study ion coupling and transport by NSS
- Project 7: Structure and kinetic mechanism of neurotransmitter transporters
- Project 8: Develop novel approaches to measure transporter-ligand interactions
- Project 9: Integrative modelling of ligand binding, transporter stability and the transport cycle
- Project 10: Time resolved structural studies of membrane
- Project 11: Single-molecule studies of secondary-active transporters and their environmental dependence
- Project 12: Resolving coarse-grained dynamic distance constraints of the transport cycle of NSS transporters using EPR spectroscopy
- Project 13: Characterization of the molecular mode of action of novel psychoactive substances
- Project 14: Development of a microfluidic cell perfusion station for controlled compound delivery and real time fluorescence monitoring
- Project 15: Assessing the molecular determinants of novel disease causing mutations in DAT
Inhaltsbereich
Project 12: Resolving coarse-grained dynamic distance constraints of the transport cycle of NSS transporters using EPR spectroscopy
Accurate structural determination of different conformational states during the transport cycle remains challenging, which is further complicated by varying degrees of conformational distribution. ESR12 will employ advanced EPR spectroscopy to derive structural information in form of distances and angles constraints for state-specific structural ensembles. The use of multiple labels and especially the integration of both NMR and EPR techniques and their application to membrane proteins is a novel and innovative aspect of this project. The specific objectives of ESR12 are:
- (i) to establish an efficient spin labelling protocol for a set of cysteine variants to study conformations and dynamics of LeuT and DAT in classically reconstituted proteoliposomes and in detergent-free encapsulated SMALPS.
- (ii) to determine coarse-grained distance restraints of the transport cycle using on DEER/PELDOR and/or RIDME distance measurements.
- (iii) to establish in close collaboration with ESR2 a combined NMR and EPR approach to determine transporter dynamics during ligand binding, occlusion and release.
- (iv) to develop a multi-label triangulation approach using iDEER and STD-NMR to measure distances and orientations.
