Developing and industrialising attachment of chemical probes to monitor biological events

This project will be completed with the CASE partner, Nanotemper Technologies GmbH. We use biophysical methods to study the architecture and functional dynamics of membrane proteins, especially many medically-relevant membrane transport systems. There is increasing evidence that membrane proteins do not act alone, but that they are organised as nano-machines which function through the concerted action of individual components with high precision and specificity observed in both time and space. We are seeking to unravel the principles underlying the architecture and dynamics of these protein nano-machines as well as their function. Our experimental approach focuses on the use of novel probes together with biophysical methods (fluorescence and magnetic resonance techniques) in combination with molecular biological, biochemical and chemical approaches.

This project will study a specific class of membrane transporter (the SLC6 sodium symport transporter LeuT), which is a structural homologue of the human neurotransmitter transporters for dopamine, serotonin, norepinephrine and amino butyric acid. These human transporters are implicated in several diseased states including depression, anxiety and attention-deficit hyperactivity disorder. Several prescribed medications target these transporters and illicit street drugs like cocaine or amphetamine also interact with them.

In addition, further high-value pharmaceutical targets such as Cereblon and Von Hippel-Lindau (VHL) proteins will also be studied. We aim to probe functional dynamics of these proteins by developing novel labelling approaches amenable to high-throughput science in combination with a range of biophysical methods.

The successful candidate will receive world class training and become competent in a broad range of biochemical and biophysical techniques working within a team environment using state-of-the-art equipment to study important families of proteins with wide ranging societal impact.

References

Tsalagradas P, Eke C, Andrews C & MacMillan F. Exploring the structural dynamics of LeuT using EPR spectroscopy: A focus on transmembrane helix 10, J. Neurochemistry in revision (2024)

Barber-Zucker S, Hall J, Froes A, Kolusheva A, MacMillan F* & Zarivach R*, The cation diffusion facilitator protein MamM’s cytoplasmic domain exhibits metal-type dependent binding modes and discriminates against Mn2+, J Biol Chem 295, 16614-16629 (2020).

Hall J, Sohail A, Cabrita EJ, Macdonald C, Stockner T, Sitte HH, Angulo J, MacMillan F, Saturation transfer difference NMR on the integral trimeric membrane transport protein GltPh determines cooperative substrate binding, Scientific Reports 10, 16483 (2020).

Bartoschik T, Gupta A, Kern B, Hitchcock A, Adams NBP, Tschammer N Quantifying the Interaction of Phosphite with ABC Transporters: MicroScale Thermophoresis and a Novel His-Tag Labeling Approach in: Postis VLG, Goldman A (Eds) Biophysics of Membrane Proteins. Methods in Molecular Biology, vol 2168. Humana, New York, NY. (2020).

Jürgens DC, Winkeljann B, Kolog Gulko M, Jin Y, Möller J, Winkeljann J, Sheshachala S, Anger A, Hörner A, Adams NBP, Urbanetz N, & Merkel OM, Efficient and Targeted siRNA Delivery to M2 Macrophages by Smart Polymer Blends for M1 Macrophage Repolarization as a Promising Strategy for Future Cancer Treatment, ACS Biomaterials Science & Engineering 10, 166-177 (2024)