Key processes of biological condensates are diffusion and material exchange with their environment. Experimentally, diffusive dynamics are typically probed via fluorescent labels. However, to date, a physics-based, quantitative framework for the dynamics of labeled condensate components is lacking. Here we derive the corresponding dynamic equations, building on the physics of phase separation, and quantitatively validate the related framework via experiments. We show that by using our framework we can precisely determine diffusion coefficients inside liquid condensates via a spatio-temporal analysis of fluorescence recovery after photobleaching (FRAP) experiments. We showcase the accuracy and precision of our approach by considering space- and time-resolved data of protein condensates and two different polyelectrolyte-coacervate systems. Interestingly, our theory can also be used to determine a relationship between the diffusion coefficient in the dilute phase and the partition coefficient, without relying on fluorescence measurements in the dilute phase. This enables us to investigate the effect of salt addition on partitioning and bypasses recently described quenching artifacts in the dense phase. Our approach opens new avenues for theoretically describing molecule dynamics in condensates, measuring concentrations based on the dynamics of fluorescence intensities, and quantifying rates of biochemical reactions in liquid condensates.
Code for modelling and data analysis is available at https://gitlab.pks.mpg.de/mesoscopic-physics-of-life/frap_theory and https://gitlab.pks.mpg.de/mesoscopic-physics-of-life/frap_analysis .
- Lars Hubatsch
- Anthony A Hyman
- Christoph A Weber
The funders had no role in study design, data collection and interpretation, or the decision to submit the work for publication.
- Rohit V Pappu, Washington University in St Louis, United States
- Received: March 21, 2021
- Accepted: October 11, 2021
- Accepted Manuscript published: October 12, 2021 (version 1)
© 2021, Hubatsch et al.
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