Single-transporter biochemistry: real-time observation of substrate transport and energy conversion of primary-active membrane transporters

Prof. Thorben Cordes, LMU Biocenter

Motivation: Membrane proteins constitute the prime target of pharmaceutical drugs and an important step in designing novel therapeutic strategies is a detailed understanding of molecular mechanisms. Single-molecule-based approaches could facilitate a next breakthrough in mechanistic understanding of transporters via (simultaneous) real-time observation of transport, energy usage and structural changes. Although there have been recent advances into this direction, mainly in the investigations of ion channels by combining optical and electrophysiological methods, similar progress has not been made for active membrane transporters. Our group has started over the past years to use single-molecule techniques for the characterization of conformational dynamics in membrane transporters[1-3], studies which you will extend towards the biochemical aspects of transport.

Project: The interdisciplinary project "Single-transporter biochemistry: real-time observation of substrate transport and energy conversion of primary-active membrane transporters" bridges state-of-the-art single-molecule fluorescence techniques, advanced data analysis and mechanistic studies of protein biochemistry. The objectives of the project are to develop assays to monitor transport and energy conversion of single membrane transporters using fluorescent sensors in combination with protein and liposome markers. This approach will provide direct access to initial transport rates, functional dynamics (on/off periods) or static heterogeneity of single transporters (see Figure).[1]

References:
[1] R. Mächtel, et al., An integrated transport mechanism of the maltose ABC importer, Research in Microbiology 170 (2019) 321-337.
[2] M. de Boer, et al., Conformational and dynamic plasticity in substrate-binding proteins underlies selective transport in ABC importers, eLife 8 (2019) e44652.
[3] F. Husada, et al., Conformational dynamics of the ABC transporter McjD seen by single‐molecule FRET, The EMBO Journal 37 (2018) e100056.
[4] G. Gouridis, et al., Conformational Dynamics in Substrate-Binding Domains Influence Transport in the ABC Importer GlnPQ“, Nature Structural and Molecular Biology 22 (2015) 57-64.

Requirements: You have a degree in Physics with a solid background in (molecular) biophysics, optics, spectroscopy, data analysis or other relevant areas. The work on this DFG-funded project will be based in the Cordes lab at the LMU Biocenter in Martinsried. During the project you will be embedded into a vibrant PhD community and other networks including the Center for NanoScience (CeNS).

Position/Salary: TV-L E13 65% - earliest entry date: 01.01.2023

Website: https://www.mikrobiologie.biologie.uni-muenchen.de/forschung/ag_cordes/index.html

For a full application send a personal motivation letter, CV, summary of research experience, study records, certificates and contact information of two referees as a single PDF to cordesbio.lmu.de. Informal inquiries via email or phone are welcome.