Prof. Simon Scheuring (Weill Cornell Medical College, US)

Title of a talk: High-Speed Atomic Force Microscopy for Dynamic Single Molecule Structural Biology

High-speed atomic force microscopy (HS-AFM) is a powerful technique that provides dynamic movies of biomolecules at work. First, I will briefly review our recent developments to break temporal limitations to characterize molecular dynamics by developing HS-AFM line scanning (HS-AFM-LS) and HS-AFM height spectroscopy (HS-AFM-HS) [1], and resolution limitations by developing Localization AFM (LAFM) [2]. Then, I will detail how we used HS-AFM to analyze membrane-embedded TRPV3 channels at the single-molecule level, and discovered a previously unobserved, transient, and reversible pentameric state in TRPV3 [3]. Finally, I will report about our most recent efforts to introduce single-molecule structural biology solving multiple structures of a single molecule along its functional timeline [4].

Figure: HS-AFM discovered a transient and reversible pentameric state in TRPV3 (top). Adaptation of protein purification and sample preparation protocols allowed to solve cryo-EM structures of the canonical tetrameric (2.6Å) and noncanonical pentameric (4.4Å) TRPV3 channel states (bottom). 

References:

[1] Heath et al., Nature Communications, 2018, 9(1):4983, High-Speed AFM Height Spectroscopy (HS-AFM-HS): Microsecond dynamics of unlabeled biomolecules.

[2] Heath et al., Nature, 2021, 594(7863):385–390, doi:10.1038/s41586-021-03551-x, Localization Atomic Force Microscopy.

[3] Lansky et al., Nature, 2023, 621(7977), 206–214, doi:10.1038/s41586-023-06470-1, A pentameric TRPV3 channel with a dilated pore.

[4] Jiang et al., Nature Struct & Mol Biol, 2024, in press, Single-Molecule Structural Biology Uncovers Basis of Transporter Wanderlust Kinetics

 

Prof. Yifan Cheng  (HHMI/University of California San Francisco, US)

Prof. Chan Cao (University of Geneva)

Prof. Sara Lin (Linköping University)