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The authors use high-speed AFM to study the interaction of yeast septin filaments with yeast lipid membranes, showing that septin is lipid phase selective and organizes into higher-order structures without the contribution of other cellular components.

An improved structure of the TRPV3 pentamer, together with molecular dynamics simulations, provides insights into its conformation, subunit interfaces, permeability to large cations, and the mechanism of transition to the canonical tetrameric state.

Bcs1, a transmembrane AAA-ATPase, facilitates the translocation of folded ISP across the inner mitochondrial membrane. This study shows that the Bcs1 ATPase cycle conformational changes are highly concerted, unlike the canonical hand-over-hand mechanism.

Here, the authors developed a pipeline to transform AFM data into 3D-density files for cross-methodology structural biology analysis. Combined with molecular dynamics flexible fitting, the approach yields structural information on protein dynamics.

Combining high-speed atomic force microscopy (AFM) with localization AFM and principal component analysis, the authors present six structures of a glutamate transporter and associate the conformational states to the molecule’s activity timeline.

Using high-speed atomic force microscopy, the authors reveal the polymerization mechanism of VIPP1 into ESCRT-III-like filaments that adopt spiral and polygonal supramolecular structures.

High-speed atomic force microscopy and molecular simulations are used to form a probability map of the interactions and motions of a protein in a lipid membrane.

Using atomic force microscopy, Pan et al. show that cyclic nucleotide-gated ion channel SthK, which can be differentially activated by cAMP and cGMP, binds both cyclic nucleotides but only cAMP can access a deep-bound state that could be essential for cAMP-dependent channel activation.

Some ion channels are modulated by binding of cyclic nucleotide to a cyclic nucleotide-binding domain. Here, the authors use high-speed atomic force microscopy to directly monitor the conformational changes induced by ligand binding to a cyclic nucleotide-modulated channel from Mesorhizobium loti.

Scanning probe microscopy techniques are hard to apply to live cell membrane imaging at high resolution as the temporal and force sensitivity are insufficient to monitor the fast processes. Colom et al.present a solution to this problem by combining high-speed atomic force microscopy with optical microscopy.

High-speed atomic force microscopy single-molecule imaging and cryo-EM analysis discover and reveal the structure of a TRPV3 pentamer, providing evidence for a non-canonical pentameric TRP-channel assembly, laying the foundation for new directions in TRP channel research.

High-Speed Atomic Force Microscopy movies of membrane proteins — diffusing and interacting in bilayers of controlled thickness — allow the determination of membrane-mediated membrane protein interaction energetics.

A high-speed atomic force microscope, which has been adapted for biological applications through the integration of a pumping system for buffer exchange and a pulsed-laser system for uncaging caged compounds, can be used to study the structure, dynamics and interaction of annexin assemblies.

Snf7 is the major component of the ESCRTIII membrane deformation system. Here, the authors used high-speed AFM to study Snf7 on nano-patterned and soft supports and show that loaded Snf7 spiral springs are curvature sensitive and deform membranes.

The molecular determinants underlying ligand gating of cyclic nucleotide-modulated ion channels remain unclear. Kowal et al.determine the conformational changes underlying cAMP binding to the bacterial channel MloK1, and propose a mechanism for coupling of ligand gating and voltage sensing in eukaryotic HCN channels.

Direct visualization of the structural transformation from pre-pore to pore of the mammalian immune defense complex perforin-2 (PFN2) reveals a clockwise hand-over-hand mechanism that propagates at ~15 subunits per second.

Septins are GTP-binding proteins involved in diverse cellular processes including division, polarity maintenance and membrane remodeling. Here authors use high-speed atomic force microscopy to show that assembly of septin filaments is a diffusion-driven process, while septin assembly into higher-order involves septin self-templating

Here, the authors use high-speed atomic force microscopy (HS-AFM) methods to characterize the single molecule kinetics of wild-type bacteriorhodopsin (bR) with millisecond temporal resolution, providing new insights into the bR conformational cycle.

Excitatory amino acid transporters (EAATs) are crucial for the removal of excitatory amino acids from the synaptic cleft. Here authors combined high-speed atomic force microscopy line-scanning with automated state assignment for the determination of transport dynamics of Glt_Ph, a prokaryotic EAAT homologue, with millisecond temporal resolution.

Cryo-electron microscopy and high-speed atomic force microscopy reveal that PIEZO1 can reversibly deform its shape towards a planar structure, which may explain how the PIEZO1 channel is gated in response to mechanical stimulation.

Annexins are cytoplasmic proteins, which bind to membranes exposing negatively charged phospholipids in a Ca²⁺-dependent manner. Here the authors use high-speed atomic force microscopy and other techniques to show that annexin-V self-assembles into highly structured lattices that lead to a membrane phase transition on PS-rich membranes.

Lipid scrambling is required for many cellular processes but the lipid-protein interactions that occur during transport are unknown. Using cryoEM and biochemical assays the authors show that membrane thinning is critical for lipid flipping.

High-speed atomic force microscopy height spectroscopy and single channel electrophysiology recordings are used to correlate conformational and functional dynamics of the model membrane protein, outer membrane protein G (OmpG). These techniques show that both states coexist and rapidly interchange in all conditions supported by molecular dynamics simulations.

The dynamics of biomolecules can occur over a wide range of time and length scales. Here the authors develop a high-speed AFM height spectroscopy method to directly detect the motion of unlabeled molecules at Angstrom spatial and microsecond temporal resolution.

A localization algorithm is applied to datasets obtained with conventional and high-speed atomic force microscopy to increase image resolution beyond the limits set by the radius of the tip used.

Cyclic nucleotide-gated (CNG) ion channels are non-selective cation channels key to signal transduction, but conformational changes associated with gating remained unknown. Here authors use high-speed atomic force microscopy to visualize SthK channels dynamics in response to cyclic nucleotides.

Mierzwa et al. show that ESCRT-III subunit turnover at the cell midbody is driven by Vps4. Rapid turnover sustains the net growth of ESCRT-III assemblies in the presence of inhibitory Vps2 and Vps24 subunits.

Microrheology of cells suggests that the dynamics of single filaments in the cytoskeleton dominate at high frequencies. This response can be used to detect differences between cell types and states — including benign and malignant cancer cells.

Bian et al. discuss the utility of different variants of scanning probe microscopy, with a focus on scanning tunnelling microscopy and atomic force microscopy. They summarize how the tools are used in the life and physical sciences.

High-speed AFM enables visualization of intrinsically disordered regions (IDRs) in TRP ion channels and shows that IDRs may mediate protein-protein interactions with adjacent TRP molecules.