Sequence Determinants of Intracellular Phase Separation by Complex Coacervation of a Disordered Protein.
Sequence Determinants of Intracellular Phase Separation by Complex Coacervation of a Disordered Protein.
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DOI：
10.1016/j.molcel.2016.05.042
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发表时间：
2016-07-07
期刊：
影响因子：
16
通讯作者：
Rosen MK
中科院分区：
文献类型：
--
作者：
Pak CW;Kosno M;Holehouse AS;Padrick SB;Mittal A;Ali R;Yunus AA;Liu DR;Pappu RV;Rosen MK
由多价和内在无序蛋白质之间的集体相互作用驱动的液-液相分离被认为介导细胞中无膜细胞器的形成。使用平行的细胞和体外试验，我们表明，Nephrin细胞内结构域（NICD），无序的蛋白质，驱动细胞内相分离通过复合凝聚，从而带负电荷的NICD与带正电荷的合作伙伴共同组装，形成富含蛋白质的致密液滴。突变显示相分离的驱动力取决于整体氨基酸组成，而不是NICD的精确序列。相反，相分离是由一个或多个区域的高负电荷密度和芳族/疏水残基分布在整个蛋白质。许多无序蛋白与NICD具有相似的序列特征，表明复合凝聚可能是促进细胞内相分离的广泛使用的机制。
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.