Project R-12619

NATIVE - Single molecule optical microscopy and lipid nanotechnology 'ménage-à-deux' for deciphering multidrug transport in near-native environments (Research)

Structural biology provides 3D architectural insights into the working of biomolecules and is key for drug design. Major challenges are functionally dynamic molecules and molecules such as membrane proteins that need a complex physiological environment to function properly. In this project we tackle these challenges by combining expertise in membrane protein biochemistry, advanced membrane nanotechnology and single-molecule fluorescence microscopy. We establish a novel and broadly applicable fluorescence imaging system based on light-sheet illumination that allows visualizing and quantifying complex structural rearrangements of membrane proteins with unprecedented ease and throughput, and most importantly, in a physiologic near-native context. We validate our developments by addressing a pressing question in the field of antibiotic resistance: How can multidrug transporters recognize and expel different molecules and antibiotics out of the cell? Practically, we first focus on a model protein for multidrug transport, L.lactis LmrP, after which we pin down the fundamental principles of multidrug transport by investigating the LmrP homologues NorA and QacA from the pathogenic S.aureus. We expect our project to cause a methodological paradigm shift in the membrane protein structural biology field and to provide opportunities for many collaborative biological investigations.

01 January 2022 - 31 December 2025