Sequence Determinants of Intracellular Phase Separation by Complex Coacervation of a Disordered Protein

Chi W. Pak, Martyna Kosno, Alex S. Holehouse, Shae B. Padrick, Anuradha Mittal, Rustam Ali, Ali A. Yunus, David R. Liu, Rohit V. Pappu, Michael K. Rosen

Research output: Contribution to journalArticle

198 Scopus citations

Abstract

Liquid-liquid phase separation, driven by collective interactions among multivalent and intrinsically disordered proteins, is thought to mediate the formation of membrane-less organelles in cells. Using parallel cellular and in vitro assays, we show that the Nephrin intracellular domain (NICD), a disordered protein, drives intracellular phase separation via complex coacervation, whereby the negatively charged NICD co-assembles with positively charged partners to form protein-rich dense liquid droplets. Mutagenesis reveals that the driving force for phase separation depends on the overall amino acid composition and not the precise sequence of NICD. Instead, phase separation is promoted by one or more regions of high negative charge density and aromatic/hydrophobic residues that are distributed across the protein. Many disordered proteins share similar sequence characteristics with NICD, suggesting that complex coacervation may be a widely used mechanism to promote intracellular phase separation.

Original languageEnglish (US)
Pages (from-to)72-85
Number of pages14
JournalMolecular cell
Volume63
Issue number1
DOIs
StatePublished - Jul 7 2016

ASJC Scopus subject areas

  • Molecular Biology
  • Cell Biology

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    Pak, C. W., Kosno, M., Holehouse, A. S., Padrick, S. B., Mittal, A., Ali, R., Yunus, A. A., Liu, D. R., Pappu, R. V., & Rosen, M. K. (2016). Sequence Determinants of Intracellular Phase Separation by Complex Coacervation of a Disordered Protein. Molecular cell, 63(1), 72-85. https://doi.org/10.1016/j.molcel.2016.05.042