Taking the heat together Many of the processes in living cells are mediated by protein complexes that dynamically assemble and dissociate depending on cellular needs. Tan et al. developed a method called thermal proximity coaggregation (TPCA) to monitor the dynamics of native protein complexes inside cells (see the Perspective by Li et al.). The method is based on the idea that proteins within a complex will coaggregate upon heat denaturation. It uses a previously described cellular shift assay to determine melting curves for thousands of proteins and assigns a TPCA signature on the basis of similarity between the curves. The method was validated by detection of many known protein complexes. It identified cell-specific interactions in six cell lines, highlighting the potential for identifying protein complexes that are modulated by disease. Science, this issue p. 1170; see also p. 1105 A readily deployable approach for system-wide intracellular studies of protein complex dynamics in nonengineered cells and tissues is discussed. Proteins differentially interact with each other across cellular states and conditions, but an efficient proteome-wide strategy to monitor them is lacking. We report the application of thermal proximity coaggregation (TPCA) for high-throughput intracellular monitoring of protein complex dynamics. Significant TPCA signatures observed among well-validated protein-protein interactions correlate positively with interaction stoichiometry and are statistically observable in more than 350 annotated human protein complexes. Using TPCA, we identified many complexes without detectable differential protein expression, including chromatin-associated complexes, modulated in S phase of the cell cycle. Comparison of six cell lines by TPCA revealed cell-specific interactions even in fundamental cellular processes. TPCA constitutes an approach for system-wide studies of protein complexes in nonengineered cells and tissues and might be used to identify protein complexes that are modulated in diseases.

中文翻译：

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热邻近共聚集用于细胞中蛋白质复合动态的系统范围分析

将热量集中起来 活细胞中的许多过程是由蛋白质复合物介导的，蛋白质复合物根据细胞需要动态组装和解离。谭等人。开发了一种称为热邻近共聚集 (TPCA) 的方法来监测细胞内天然蛋白质复合物的动态（参见 Li 等人的观点）。该方法基于复合物中的蛋白质在热变性时会凝聚的想法。它使用先前描述的细胞移位分析来确定数千种蛋白质的熔解曲线，并根据曲线之间的相似性分配 TPCA 特征。该方法通过检测许多已知的蛋白质复合物进行了验证。它鉴定了六种细胞系中的细胞特异性相互作用，突出了鉴定受疾病调节的蛋白质复合物的潜力。科学，这个问题 p。1170; 另见第。1105 讨论了一种易于部署的方法，用于对非工程细胞和组织中的蛋白质复合物动力学进行系统范围的细胞内研究。蛋白质在不同细胞状态和条件下彼此之间存在差异性相互作用，但缺乏有效的全蛋白质组策略来监测它们。我们报告了热邻近共聚（TPCA）在蛋白质复合物动力学的高通量细胞内监测中的应用。在经过充分验证的蛋白质-蛋白质相互作用中观察到的显着 TPCA 特征与相互作用的化学计量呈正相关，并且在超过 350 种注释的人类蛋白质复合物中可从统计学上观察到。使用 TPCA，我们鉴定了许多未检测到差异蛋白表达的复合物，包括染色质相关复合物，在细胞周期的 S 期进行调节。TPCA 对六种细胞系的比较揭示了细胞特异性相互作用，即使在基本细胞过程中也是如此。TPCA 构成了对非工程细胞和组织中蛋白质复合物进行全系统研究的方法，可用于鉴定在疾病中调节的蛋白质复合物。