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Single-droplet surface-enhanced Raman scattering decodes the molecular determinants of liquid-liquid phase separation

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  • Anamika Avni

    (Indian Institute of Science Education and Research (IISER) Mohali
    Indian Institute of Science Education and Research (IISER) Mohali)

  • Ashish Joshi

    (Indian Institute of Science Education and Research (IISER) Mohali
    Indian Institute of Science Education and Research (IISER) Mohali)

  • Anuja Walimbe

    (Indian Institute of Science Education and Research (IISER) Mohali
    Indian Institute of Science Education and Research (IISER) Mohali)

  • Swastik G. Pattanashetty

    (Indian Institute of Science Education and Research (IISER) Mohali
    Indian Institute of Science Education and Research (IISER) Mohali)

  • Samrat Mukhopadhyay

    (Indian Institute of Science Education and Research (IISER) Mohali
    Indian Institute of Science Education and Research (IISER) Mohali
    Indian Institute of Science Education and Research (IISER) Mohali)

Abstract

Biomolecular condensates formed via liquid-liquid phase separation (LLPS) are involved in a myriad of critical cellular functions and debilitating neurodegenerative diseases. Elucidating the role of intrinsic disorder and conformational heterogeneity of intrinsically disordered proteins/regions (IDPs/IDRs) in these phase-separated membrane-less organelles is crucial to understanding the mechanism of formation and regulation of biomolecular condensates. Here we introduce a unique single-droplet surface-enhanced Raman scattering (SERS) methodology that utilizes surface-engineered, plasmonic, metal nanoparticles to unveil the inner workings of mesoscopic liquid droplets of Fused in Sarcoma (FUS) in the absence and presence of RNA. These highly sensitive measurements offer unprecedented sensitivity to capture the crucial interactions, conformational heterogeneity, and structural distributions within the condensed phase in a droplet-by-droplet manner. Such an ultra-sensitive single-droplet vibrational methodology can serve as a potent tool to decipher the key molecular drivers of biological phase transitions of a wide range of biomolecular condensates involved in physiology and disease.

Suggested Citation

  • Anamika Avni & Ashish Joshi & Anuja Walimbe & Swastik G. Pattanashetty & Samrat Mukhopadhyay, 2022. "Single-droplet surface-enhanced Raman scattering decodes the molecular determinants of liquid-liquid phase separation," Nature Communications, Nature, vol. 13(1), pages 1-13, December.
    • Handle: RePEc:nat:natcom:v:13:y:2022:i:1:d:10.1038_s41467-022-32143-0
    DOI: 10.1038/s41467-022-32143-0
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    1. Joshua A. Riback & Lian Zhu & Mylene C. Ferrolino & Michele Tolbert & Diana M. Mitrea & David W. Sanders & Ming-Tzo Wei & Richard W. Kriwacki & Clifford P. Brangwynne, 2020. "Composition-dependent thermodynamics of intracellular phase separation," Nature, Nature, vol. 581(7807), pages 209-214, May.
    2. Taranpreet Kaur & Muralikrishna Raju & Ibraheem Alshareedah & Richoo B. Davis & Davit A. Potoyan & Priya R. Banerjee, 2021. "Sequence-encoded and composition-dependent protein-RNA interactions control multiphasic condensate morphologies," Nature Communications, Nature, vol. 12(1), pages 1-16, December.
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    Cited by:

    1. Saumyak Mukherjee & Lars V. Schäfer, 2023. "Thermodynamic forces from protein and water govern condensate formation of an intrinsically disordered protein domain," Nature Communications, Nature, vol. 14(1), pages 1-13, December.
    2. Ashish Joshi & Anuja Walimbe & Anamika Avni & Sandeep K. Rai & Lisha Arora & Snehasis Sarkar & Samrat Mukhopadhyay, 2023. "Single-molecule FRET unmasks structural subpopulations and crucial molecular events during FUS low-complexity domain phase separation," Nature Communications, Nature, vol. 14(1), pages 1-17, December.
    3. Agustín Mangiarotti & Macarena Siri & Nicky W. Tam & Ziliang Zhao & Leonel Malacrida & Rumiana Dimova, 2023. "Biomolecular condensates modulate membrane lipid packing and hydration," Nature Communications, Nature, vol. 14(1), pages 1-19, December.

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