Unraveling Molecular Dynamics
Acknowledgements We would like to thank Carmen M. Domínguez and
Christof M. Niemeyer (KIT, Karlsruhe, Germany) for provid- ing the DON samples used in Figure 2.
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Figure 5: High-speed AFM imaging of an A5 P6 honeycomb lattice, with cen- tral non-P6 positions occupied by mobile A5 trimers (top left). A 3D view of a “2IE7” A5 PDB trimer (from
www.rcsb.org) with a global C3 symmetry (top right) is given for comparison only. Outlined inset is shown in the bottom panel at tem- poral resolution of 833 ms, identifying several preferred orientations of the rota- tional A5 dynamics. XYZ-scales in top left and bottom panels are 100×100×1.5 nm3
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in combination with the high temporal resolution, here can be applied as a molecular fingerprinting tool for studying collagen type I mutations in lab diagnostics. The high-speed AFM imaging of mobile A5 trimers
within the stationary P6 lattice indicate that preferred structural orientations at 60° can be studied (Figure 5), but realistically the rotational dynamics of the process is at least a few orders of magnitude faster than the high-speed AFM imaging rates. As suggested previously [24], further studies of A5 rotation kinetics would require the application of single line, or even point scanning, to boost the temporal resolution.
Conclusion Tis article describes the application of high-speed AFM
for studying several biological systems with acquisition rates of up to 50 frames/sec. With no requirement for sample pro- cessing, high-speed AFM enables measurements of samples at their near-native state. With acquisition line rates of up to 5 kHz, the high-speed AFM used here offers a 3-fold boost in temporal resolution compared to conventional AFM. In turn, this enables the real-time studies of dynamic and molecular interactions such as single molecule binding dynamics, track- ing of protein-protein and protein-DNA interactions, DNA rehybridization dynamics, monitoring of enzyme kinetics, lipid remodeling in multi-component membranes, etc. Cur- rent studies are underway to demonstrate how high-speed force spectroscopy applications, including nanomechanical mapping of single molecules, membrane segregation, and novel unfolding pathways of biomolecules in health and dis- ease would additionally complement our knowledge of the molecular dynamics in both life science and material science applications.
14
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www.microscopy-today.com • 2022 May J 104 (2013) https://doi
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