The self-assembly of proteins has been widely studied in controlled in vitro conditions, and more recently in biological environments. The self-assembly of proteins in biology can be a feature of the pathogenesis of protein condensation disease, or can occur during normal physiological function, for example during the formation of intracellular non-membrane bound organelles. To determine the mechanisms for the assembly process fully, controlled in vitro experiments using purified protein solutions are often required. However, making direct connections between insights gathered from controlled experiments and those in complex biological environments remains a challenge. Using the P23T mutant of human γD-crystallin, a protein associated with congenital cataract, we have demonstrated that the equilibrium solubility boundary and solution behavior measured using phase diagrams of purified protein solutions is consistent with the assembly of the protein expressed in cell-free expression medium in artificial cells (without fluorescent labelling) and condensates formed in mammalian cells, thereby directly connecting in vitro measurements with those performed under physiological conditions.

中文翻译：

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在人工和哺乳动物细胞中原位表达后的蛋白质自组装。

蛋白质的自组装已在受控的体外条件下进行了广泛研究，最近在生物环境中也进行了研究。生物学中蛋白质的自组装可能是蛋白质凝结疾病发病机制的一个特征，或者可以发生在正常的生理功能过程中，例如在细胞内非膜结合细胞器的形成过程中。为了充分确定组装过程的机制，通常需要使用纯化的蛋白质溶液进行受控的体外实验。然而，在受控实验收集的见解与复杂生物环境中的见解之间建立直接联系仍然是一个挑战。使用人类γD-晶状体蛋白（一种与先天性白内障相关的蛋白质）的P23T突变体，