Mapping Phase Diagram of Tau-RNA LLPS Under Live Cell Coculturing Conditions

Yanxian Lin; Yann Fichou; Jennifer N Rauch; Xuemei Zhang; Kenneth S Kosik; Songi Han

doi:10.1007/978-1-0716-2597-2_18

Mapping Phase Diagram of Tau-RNA LLPS Under Live Cell Coculturing Conditions

Methods Mol Biol. 2023:2551:269-284. doi: 10.1007/978-1-0716-2597-2_18.

Authors

Yanxian Lin¹, Yann Fichou², Jennifer N Rauch^{3

4}, Xuemei Zhang⁴, Kenneth S Kosik^{3

4}, Songi Han^{5

6}

Affiliations

¹ Biomolecular Science and Engineering, University of California, Santa Barbara, CA, USA.
² Department of Chemistry and Biochemistry, University of California, Santa Barbara, CA, USA.
³ Molecular, Cell and Developmental Biology, University of California, Santa Barbara, CA, USA.
⁴ Neuroscience Research Institute, University of California, Santa Barbara, CA, USA.
⁵ Department of Chemistry and Biochemistry, University of California, Santa Barbara, CA, USA. songihan@ucsb.edu.
⁶ Department of Chemical Engineering, University of California, Santa Barbara, CA, USA. songihan@ucsb.edu.

PMID: 36310209
DOI: 10.1007/978-1-0716-2597-2_18

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.

Keywords: Complex coacervation; Double electron-electron resonance; Liquid-liquid phase separation; Phase diagram; Tau protein.

Publication types

Research Support, Non-U.S. Gov't
Research Support, N.I.H., Extramural

MeSH terms

RNA* / metabolism

Substances

RNA