Project description

Building on our recent work enabling the single-molecule measurement of transmembrane transport proteins in lipid nanodiscs using optical tweezers [1], this project aims to expand our understanding of how the physical and chemical properties of the environment impact the stability and function of archaeal and human secondary active transporters.

Objective: We seek to elucidate the timescales and environmental influences on the structural rearrangements of secondary active transporters, such as GltTk [2], [3], which are crucial for cellular homeostasis.

Significance: Transmembrane transport proteins are essential for regulating the uptake of vital molecules that cells cannot produce themselves. Understanding their functional dynamics in different environments can provide insights into cellular regulation and potential therapeutic targets.

Methodology: Using optical tweezers, we will perform real-time, nanoscale measurements of the structural rearrangements of archaeal and human transporters. This technique allows us to capture the kinetic and structural changes with high precision, making it ideal for our study.

Expected Outcomes: We anticipate gaining an in-depth understanding of the differences in regulatory processes between archaeal and human transporters, shedding light on their evolutionary adaptations and functional mechanisms.

Bibliography:

[1] L. van der Sleen, J. A. Stevens, S. J. Marrink, B. Poolman, and K. Tych, “Probing the stability and interdomain interactions in the ABC transporter OpuA using single-molecule optical tweezers,” Cell Rep, vol. 43, no. 4, p. 114110, Apr. 2024, doi: 10.1016/j.celrep.2024.114110.

[2] S. Jensen, A. Guskov, S. Rempel, I. Hänelt, and D. J. Slotboom, “Crystal structure of a substrate-free aspartate transporter,” Nat Struct Mol Biol, vol. 20, no. 10, pp. 1224–1226, Oct. 2013, doi: 10.1038/nsmb.2663.

[3] G. Trinco et al., “Kinetic mechanism of Na+-coupled aspartate transport catalyzed by GltTk,” Commun Biol, vol. 4, no. 1, pp. 1–11, Jun. 2021, doi: 10.1038/s42003-021-02267-y.

Additional specifications

We are seeking candidates with backgrounds in molecular biology, biochemistry or biophysics with experience in protein expression and purification protocols. Prior experience with biophysical methods, optical tweezers and/or with membrane proteins is not essential, but is highly relevant. Full training will be provided.