@article{72e4f52f376c4c018146281febf5cab7,
title = "Sequence Determinants of Intracellular Phase Separation by Complex Coacervation of a Disordered Protein",
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.",
author = "Pak, {Chi W.} and Martyna Kosno and Holehouse, {Alex S.} and Padrick, {Shae B.} and Anuradha Mittal and Rustam Ali and Yunus, {Ali A.} and Liu, {David R.} and Pappu, {Rohit V.} and Rosen, {Michael K.}",
note = "Funding Information: We thank Thomas Scheuermann for help with CD experiments and analysis; Kate Luby-Phelps and Abhijit Bugde in the UTSW Live Cell Imaging Facility for assistance with microscopy; Shinichi Nakagawa for sharing NEAT1 MEFs; Nina Jones and Tony Pawson for sharing Nephrin expression constructs; and Tyler Harmon for discussion. Research was supported by the Howard Hughes Medical Institute (M.K.R. and D.R.L.), grants from the NIH (R01-GM56322, M.K.R.; R01-NS056114, R.V.P.; R01-GM095501, D.R.L.; and F32-DK091074, C.W.P.), and the Welch Foundation (I–1544, M.K.R.). The UTSW Live Cell Imaging Facility is a Shared Resource of the Harold C. Simmons Cancer Center and is supported in part by NCI Cancer Center Support Grant 1P30 CA142543-01. D.R.L. is the founder of Permeon Biologics, a company that applies supercharged proteins for macromolecular delivery. Publisher Copyright: {\textcopyright} 2016 Elsevier Inc.",
year = "2016",
month = jul,
day = "7",
doi = "10.1016/j.molcel.2016.05.042",
language = "English (US)",
volume = "63",
pages = "72--85",
journal = "Molecular Cell",
issn = "1097-2765",
publisher = "Cell Press",
number = "1",
}