scholarly journals Quantification of Biomolecular Dynamics Inside Real and Synthetic Nuclear Pore Complexes Using Time-Resolved Atomic Force Microscopy

ACS Nano ◽  
2019 ◽  
Vol 13 (7) ◽  
pp. 7949-7956 ◽  
Author(s):  
George J. Stanley ◽  
Bernice Akpinar ◽  
Qi Shen ◽  
Patrick D. Ellis Fisher ◽  
C. Patrick Lusk ◽  
...  
2010 ◽  
Vol 171 (2) ◽  
pp. 154-162 ◽  
Author(s):  
Ning-Ping Huang ◽  
Mike Stubenrauch ◽  
Joachim Köser ◽  
Nicole Taschner ◽  
Ueli Aebi ◽  
...  

2019 ◽  
Author(s):  
George J. Stanley ◽  
Bernice Akpinar ◽  
Qi Shen ◽  
Patrick D. Ellis Fisher ◽  
C. Patrick Lusk ◽  
...  

AbstractOver the past decades, atomic force microscopy (AFM) has emerged as an increasingly powerful tool to study the dynamics of biomolecules at nanometre length scales. However, the more stochastic the nature of such biomolecular dynamics, the harder it becomes to distinguish them from AFM measurement noise. Rapid, stochastic dynamics are inherent to biological systems comprising intrinsically disordered proteins. One role of such proteins is in the formation of the transport barrier of the nuclear pore complex (NPC): the selective gateway for macromolecular traffic entering or exiting the nucleus. Here, we use AFM to observe the dynamics of intrinsically disordered proteins from two systems: the transport barrier of native NPCs, and the transport barrier of a mimetic NPC made using a DNA origami scaffold. Analysing data recorded with 50-200 ms temporal resolution, we highlight the importance of drift correction and appropriate baseline measurements in such experiments. In addition, we describe an auto-correlation analysis to quantify time scales of observed dynamics and to assess their veracity — an analysis protocol that lends itself to the quantification of stochastic fluctuations in other biomolecular systems. The results reveal the surprisingly slow rate of stochastic, collective transitions inside mimetic NPCs, highlighting the importance of FG-nup cohesive interactions.


2019 ◽  
Vol 10 ◽  
pp. 617-633 ◽  
Author(s):  
Aaron Mascaro ◽  
Yoichi Miyahara ◽  
Tyler Enright ◽  
Omur E Dagdeviren ◽  
Peter Grütter

Recently, there have been a number of variations of electrostatic force microscopy (EFM) that allow for the measurement of time-varying forces arising from phenomena such as ion transport in battery materials or charge separation in photovoltaic systems. These forces reveal information about dynamic processes happening over nanometer length scales due to the nanometer-sized probe tips used in atomic force microscopy. Here, we review in detail several time-resolved EFM techniques based on non-contact atomic force microscopy, elaborating on their specific limitations and challenges. We also introduce a new experimental technique that can resolve time-varying signals well below the oscillation period of the cantilever and compare and contrast it with those previously established.


2004 ◽  
pp. 749-756
Author(s):  
R. W. Stark ◽  
G. Schitter ◽  
M. Stark ◽  
R. Guckenberger ◽  
A. Stemmer

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