Development of functional synthetic biomolecular condensates.

项目总结 无膜细胞器，或生物分子凝聚体，已经成为组织内容的一种策略 原核细胞和真核细胞。这些相分离的隔室在一系列细胞 功能--从信号到调节新陈代谢途径或控制基因表达--然而仍然有 关于它们的形成、动力和功能的基本机制的问题。两国关系 凝析油分子性质和功能之间的关系尚不清楚，但这可能提供一条途径 治疗涉及蛋白质凝聚体失调的疾病(神经变性、白内障、癌症)。我们的 研究建议开发功能性合成生物分子缩合物，以解决几个过度 探索性问题：特定的分子间相互作用(静电、阳离子-π等)如何对蛋白质有贡献 相变？蛋白质序列和结构如何影响细胞的物理性质和功能 凝聚相？材料的性质和生物分子的功能之间有联系吗 凝析油？半合成的生物分子凝聚物将使我们能够评估分子相互作用如何在 凝聚相不仅对相的动力学有贡献，而且对小分子和大分子也有贡献。 分配，最终是内源生物分子凝聚物的功能。建议的目标是 研究计划是在体外和体内创造具有酶活性的合成无膜细胞器。新的 将制备具有不同化学环境的材料和新的蛋白质凝聚物成像方法 在分子尺度上将被建立。这些进展将有助于我们理解蛋白质序列是如何 在微观尺度(例如，单个酶)和中尺度(例如，浓缩的酶)上影响功能 相细胞隔室)。工程上正交的生物分子凝聚物有可能影响我们的 了解天然生物分子缩合物的功能并提供合成生物学平台 人为地调节细胞内的信息流。

PROJECT SUMMARY Membraneless organelles, or biomolecular condensates, have emerged as a strategy to organize the contents of prokaryotic and eukaryotic cells. These phase separated compartments play key roles in a range of cellular functions – from signaling to tuning metabolic pathways or controlling gene expression – yet there are still questions about the fundamental mechanisms for their formation, dynamics, and function. The relationship between condensate molecular properties and function is not yet understood, but this could provide an avenue to treat diseases that involve dysregulated protein condensates (neurodegeneration, cataracts, cancer). Our research proposes to develop functional synthetic biomolecular condensates in order to address several over- arching questions: How do specific intermolecular interactions (electrostatic, cation-π, etc.) contribute to protein phase transitions? How do protein sequence and structure influence the physical properties and function of the condensed phase? Is there a connection between the materials properties and the function of biomolecular condensates? Semi-synthetic biomolecular condensates will allow us to evaluate how molecular interactions in the condensed phase contribute not only to the dynamics of the phase but also to small and macromolecule partitioning, and ultimately the function of endogenous biomolecular condensates. The goals of the proposed research program are to create enzymatically active synthetic membraneless organelles in vitro and in vivo. New materials with varied chemical environments will be prepared and new methods for imaging protein condensates at the molecular scale will be established. These advances will help us to understand how protein sequence influences function at both the microscale (e.g. of an individual enzyme) and the mesoscale (e.g. of a condensed phase cellular compartment). Engineering orthogonal biomolecular condensates has the potential to impact our understanding of the function of native biomolecular condensates and provide a synthetic biology platform to artificially regulate information flow in the cell.