Department of Biological Sciences, Columbia University, New York, New York, USA.
Proteomics, Max Planck Institute of Biophysics, Frankfurt am Main, Germany; Proteomics, Max Planck Institute for Brain Research, Frankfurt am Main, Germany.
Mol Cell Proteomics. 2021;20:100016. doi: 10.1074/mcp.R120.002190. Epub 2020 Dec 7.
In all cells, proteins are continuously synthesized and degraded to maintain protein homeostasis and modify gene expression levels in response to stimuli. Collectively, the processes of protein synthesis and degradation are referred to as protein turnover. At a steady state, protein turnover is constant to maintain protein homeostasis, but in dynamic responses, proteins change their rates of synthesis and degradation to adjust their proteomes to internal or external stimuli. Thus, probing the kinetics and dynamics of protein turnover lends insight into how cells regulate essential processes such as growth, differentiation, and stress response. Here, we outline historical and current approaches to measuring the kinetics of protein turnover on a proteome-wide scale in both steady-state and dynamic systems, with an emphasis on metabolic tracing using stable isotope-labeled amino acids. We highlight important considerations for designing proteome turnover experiments, key biological findings regarding the conserved principles of proteome turnover regulation, and future perspectives for both technological and biological investigation.
在所有细胞中，蛋白质不断合成和降解，以维持蛋白质的内稳态，并根据刺激来调节基因表达水平。蛋白质的合成和降解过程统称为蛋白质周转。在稳定状态下，蛋白质周转是恒定的，以维持蛋白质的内稳态，但在动态响应中，蛋白质会改变其合成和降解的速率，以调整其蛋白质组来适应内部或外部刺激。因此，探测蛋白质周转的动力学和动态可以深入了解细胞如何调节诸如生长、分化和应激反应等基本过程。在这里，我们概述了在稳态和动态系统中在全蛋白质组范围内测量蛋白质周转动力学的历史和当前方法，重点介绍了使用稳定同位素标记的氨基酸进行代谢追踪的方法。我们强调了设计蛋白质组周转实验时的重要考虑因素、关于蛋白质组周转调节的保守原则的关键生物学发现，以及技术和生物学研究的未来展望。
