Phase separation of intrinsically disordered proteins (IDPs) is a remarkable feature of living cells to dynamically control intracellular partitioning. Despite the numerous new IDPs that have been identified, progress towards rational engineering in cells has been limited. To address this limitation, we systematically scanned the sequence space of native IDPs and designed artificial IDPs (A-IDPs) with different molecular weights and aromatic content, which exhibit variable condensate saturation concentrations and temperature cloud points in vitro and in cells. We created A-IDP puncta using these simple principles, which are capable of sequestering an enzyme and whose catalytic efficiency can be manipulated by the molecular weight of the A-IDP. These results provide a robust engineered platform for creating puncta with new, phase-separation-mediated control of biological function in living cells.

内在无序蛋白质 (IDP) 的相分离是活细胞动态控制细胞内分配的显着特征。尽管已经确定了许多新的 IDP，但在细胞中进行合理工程的进展是有限的。为了解决这一限制，我们系统地扫描了天然 IDPs 的序列空间，并设计了具有不同分子量和芳烃含量的人工 IDPs (A-IDPs)，它们在体外和细胞中表现出可变的冷凝物饱和浓度和温度浊点。我们使用这些简单的原理创建了 A-IDP puncta，它们能够隔离酶，并且其催化效率可以通过 A-IDP 的分子量来控制。这些结果提供了一个强大的工程平台，用于创建具有新的 puncta，