Tethered Particle Motion Technique in Crowded Media: Compaction of DNA by Globular Macromolecules.

Tethered Particle Motion (TPM) is a single molecule technique, which consists in tracking the motion of a nanoparticle (NP) immersed in a fluid and tethered to a glass surface by a DNA molecule. The present work addresses the question of the applicability of TPM to fluids which contain crowders at volume fractions ranging from that of the nucleoid of living bacteria (around 30%) up to the jamming threshold (around 66%). In particular, we were interested in determining whether TPM can be used to characterize the compaction of DNA by globular crowders. To this end, extensive Brownian Dynamics simulations were performed with a specifically built coarse-grained model. Analysis of the simulations reveals several effects not observed in dilute media, which impose constraints on the TPM setup. In particular, the Tethered Fluorophore Motion (TFM) technique, which consists in replacing the NP by a much smaller fluorophore, is probably better suited than standard TPM. Moreover, a sample preparation technique which does not involve hydrophilic patches may be required. Finally, the use of a DNA brush may be needed to achieve DNA concentrations close to in vivo ones.

Citation

Marc Joyeux. Tethered Particle Motion Technique in Crowded Media: Compaction of DNA by Globular Macromolecules. The journal of physical chemistry. B. 2024 Jul 25;128(29):7227-7236

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PMID: 38986040

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