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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 5: Electrophysiological characterization of neurotransmitter transporters and formulation of an overall kinetic model
Our aim is the electrophysiological characterization of LeuT, DAT and GABA transporters using Solid Supported Membrane (SSM) -based electrophysiology. Our SURFE2R technology allows for real-time measurements of charge movements in the transmembrane electric field, originating from substrate translocation or electrogenic conformational transitions.
The three main objectives are:
- to develop an assay to enhance sensitivity and time resolution using SSM-based electrophysiology to increase signal-to-noise ratio for transport assays. This includes variations of the sensor surface and application of membrane potentials by chemical means.
- to electrophysiologically characterise NSS function using the SSM-based electrophysiology approach (EC50, IC50, Vmax, cooperativity).
- to develop an overall kinetic transport model by identifying and characterizing Na+, substrate and inhibitor induced electrogenic partial reactions of the transport cycle. Key is comparison of wild type transporters and transport deficient mutants as well as comparison the results obtained within NeuroTrans. Functional symmetry using different sample preparations also will be investigated.
There will be secondments with the research group of Christine Ziegler (University of Regensburg) and Thorben Cordes (University of Munich) which involve transporter assays using radioactivity labelled substrates as well as fluorescence techniques.
