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.