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Liquid-Liquid Phase Separation to Study the Association of Proteins in Solution

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Protein Aggregation

Part of the book series: Methods in Molecular Biology ((MIMB,volume 2551))

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Abstract

Liquid-liquid phase separation (LLPS) is a reversible biological process that contributes to the formation of critical concentration of proteins, forming membraneless compartments that are physiologically and pathologically relevant. Several proteins have been shown to demix into liquid droplets under in vitro crowding conditions. These studies are mainly conducted in isolation using purified recombinant proteins. Recently, we used LLPS to study the association between two proteins that are co-aggregated in Alzheimer’s disease brain, tau, and EFhd2. Here, we describe how we used LLPS to determine the molecular components that contribute to the transition of these two proteins from liquid droplets to solid-like structures.

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References

  1. Aguzzi A, Altmeyer M (2016) Phase separation: linking cellular compartmentalization to disease. Trends Cell Biol 26:547–558

    Article  CAS  PubMed  Google Scholar 

  2. Banani SF, Lee HO, Hyman AA et al (2017) Biomolecular condensates: organizers of cellular biochemistry. Nat Rev Mol Cell Biol 18:285–298

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  3. Shin Y, Brangwynne CP (2017) Liquid phase condensation in cell physiology and disease. Science 357(6357):eaaf4382

    Article  PubMed  Google Scholar 

  4. Babinchak WM, Surewicz WK (2020) Liquid-liquid phase separation and its mechanistic role in pathological protein aggregation. J Mol Biol 432:1910–1925

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  5. Ambadipudi S, Biernat J, Riedel D et al (2017) Liquid-liquid phase separation of the microtubule-binding repeats of the Alzheimer-related protein Tau. Nat Commun 8:275

    Article  PubMed  PubMed Central  Google Scholar 

  6. Hernández-Vega A, Braun M, Scharrel L et al (2017) Local nucleation of microtubule bundles through tubulin concentration into a condensed Tau phase. Cell Rep 20:2304–2312

    Article  PubMed  PubMed Central  Google Scholar 

  7. Vega IE, Traverso EE, Ferrer-Acosta Y et al (2008) A novel calcium-binding protein is associated with tau proteins in tauopathy. J Neurochem 106:96–106

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  8. Ferrer-Acosta Y, Rodríguez-Cruz EN, Orange F et al (2013) EFhd2 is a novel amyloid protein associated with pathological tau in Alzheimer’s disease. J. Neurochem 125:921–931

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  9. Vega IE, Umstead A, Kanaan NM (2019) EFhd2 affects tau liquid-liquid phase separation. Front Neurosci 13:845

    Article  PubMed  PubMed Central  Google Scholar 

  10. Combs B, Tiernan CT, Hamel C et al (2017) Production of recombinant tau oligomers in vitro. Methods Cell Biol 141:45–64

    Article  CAS  PubMed  PubMed Central  Google Scholar 

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Acknowledgment

This work was supported by institutional funds from the Department of Translational Neuroscience and the College of Human Medicine at Michigan State University.

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Correspondence to Irving E. Vega .

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Vega, I.E., Umstead, A. (2023). Liquid-Liquid Phase Separation to Study the Association of Proteins in Solution. 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_17

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  • DOI: https://doi.org/10.1007/978-1-0716-2597-2_17

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  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-0716-2596-5

  • Online ISBN: 978-1-0716-2597-2

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