Abstract
Protein liquid-liquid phase separation (LLPS) has been associated with biological functions and pathological aggregation. Mapping the phase separation conditions is the first step to identify and quantify the driving forces of LLPS. Here, we describe the protocols to draw the phase diagram of tau-RNA LLPS and use the mapped diagram to guide experimental conditions for LLPS-cell coculturing, electron resonance spectroscopy in particular double electron-electron resonance spectroscopy, crosslinking immunoprecipitation, and isothermal titration calorimetry.
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Acknowledgment
The authors acknowledge support for this work by the National Institute on Aging (NIA) of the National Institute of Health (NIH) through Grant No. RO1AG05605. KSK and SH also acknowledge partial support by the Tau Consortium of the Rainwater Foundation. We acknowledge the use of the NRI-MCDB Microscopy Facility at UC, Santa Barbara. All the funders above had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.
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Lin, Y., Fichou, Y., Rauch, J.N., Zhang, X., Kosik, K.S., Han, S. (2023). Mapping Phase Diagram of Tau-RNA LLPS Under Live Cell Coculturing Conditions. In: Cieplak, A.S. (eds) Protein Aggregation. Methods in Molecular Biology, vol 2551. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-2597-2_18
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DOI: https://doi.org/10.1007/978-1-0716-2597-2_18
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