Rxivist logo

Intramolecular dynamics of single molecules in free diffusion

By Charles Limouse, Jason C. Bell, Colin J. Fuller, Aaron F. Straight, Hideo Mabuchi

Posted 24 Mar 2017
bioRxiv DOI: 10.1101/120311

Biomolecular systems such as multiprotein complexes or biopolymers can span several tens to several hundreds of nanometers, but the dynamics of such mesocale molecules remain challenging to probe. We have developed a single-molecule technique that uses Tracking Fluorescence Correlation Spectroscopy (tFCS) to measure the conformation and dynamics of molecular assemblies specifically at the mesoscale level (100-1000 nm). tFCS is non-perturbative, as molecules, which are tracked in real-time, are untethered and freely diffusing. To achieve sub-diffraction spatial resolution, we use a feedback scheme which allows us to maintain the molecule at an optimal position within the laser intensity gradient. We find that tFCS is sufficiently sensitive to measure the distance fluctuations between two sites within a DNA molecule separated by distances as short as 1000 bp. We demonstrate that tFCS detects changes in the compaction of reconstituted chromatin, and can assay transient protein mediated interactions between distant sites in an individual DNA molecule. Our measurements highlight the impact that tFCS can have in the study of a wide variety of biochemical processes involving mesoscale conformational dynamics.

Download data

  • Downloaded 384 times
  • Download rankings, all-time:
    • Site-wide: 41,899 out of 88,415
    • In biophysics: 1,679 out of 3,846
  • Year to date:
    • Site-wide: 82,742 out of 88,415
  • Since beginning of last month:
    • Site-wide: 72,121 out of 88,415

Altmetric data


Downloads over time

Distribution of downloads per paper, site-wide


PanLingua

Sign up for the Rxivist weekly newsletter! (Click here for more details.)


News