Integrated changes in thermal stability and proteome abundance during altered nutrient states in Escherichia coli and human cells.
Integrated changes in thermal stability and proteome abundance during altered nutrient states in Escherichia coli and human cells.
复制标题
在大肠杆菌和人类细胞的营养状态改变期间，热稳定性和蛋白质组丰度的综合变化。
DOI：
10.1002/pmic.202100254
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发表时间：
2022
期刊：
影响因子：
3.4
通讯作者：
Murphy,JohnPatrick
中科院分区：
文献类型：
--
作者：
Sultonova,Mukhayyo;Blackmore,Beau;Du,Ronnie;Philips,Olivier;Paulo,JoaoA;Murphy,JohnPatrick
改变的热溶解度测量技术正在成为评估配体结合、翻译后修饰、蛋白质-蛋白质相互作用以及在不同细胞条件下影响蛋白质状态的许多其他细胞过程的强大工具。通过使用等压标记试剂，可以在全球蛋白质组规模上实现热溶解性或稳定性分析技术，这些试剂促进了测量蛋白质组跨热梯度变化所需的多路复用能力。其中的关键是热蛋白质组学(TPP)，它需要每个梯度8-10个等压标签，并生成多个蛋白质组数据集来测量不同的重复和条件。此外，使用TPP测量不同条件下的蛋白质热稳定性状态可能还需要测量不同的蛋白质丰度。在这里，我们使用蛋白质组整体稳定性改变(PISA)分析，TPP的高通量版本，来测量与其蛋白质丰度标准化的蛋白质热稳定性的全球变化。我们探索使用这种方法来确定对数期和稳定期之间的蛋白质状态的变化-大肠杆菌以及葡萄糖饥饿的人Hek293T细胞。我们观察到290和350个蛋白质中的蛋白质强度校正后的PISA变化是由于静止相转变碱引起的。结肠和葡萄糖饥饿。这些数据揭示了几个蛋白质的例子，这些蛋白质以前并不是单凭丰度就与营养状态相关的。其中包括。结肠蛋白，如推测的酰辅酶A脱氢酶(AidB)和伴侣蛋白(CnoX)，以及人RAB囊泡运输蛋白和许多其他可能表明它们参与了代谢性疾病，如癌症。
Altered thermal solubility measurement techniques are emerging as powerful tools to assess ligand binding, post‐translational modification, protein–protein interactions, and many other cellular processes that affect protein state under various cellular conditions. Thermal solubility or stability profiling techniques are enabled on a global proteomic scale by employing isobaric tagging reagents that facilitate multiplexing capacity required to measure changes in the proteome across thermal gradients. Key among these is thermal proteomic profiling (TPP), which requires 8–10 isobaric tags per gradient and generation of multiple proteomic datasets to measure different replicates and conditions. Furthermore, using TPP to measure protein thermal stability state across different conditions may also require measurements of differential protein abundance. Here, we use the proteome integral stability alteration (PISA) assay, a higher throughput version of TPP, to measure global changes in protein thermal stability normalized to their protein abundance. We explore the use of this approach to determine changes in protein state between logarithmic and stationary phaseEscherichia colias well as glucose‐starved human Hek293T cells. We observed protein intensity‐corrected PISA changes in 290 and 350 proteins due to stationary phase transition inE. coliand glucose starvation, respectively. These data reveal several examples of proteins that were not previously associated with nutrient states by abundance alone. These includeE. coliproteins such as putative acyl‐CoA dehydrogenase (aidB) and chaperedoxin (cnoX) as well as human RAB vesicle trafficking proteins and many others which may indicate their involvement in metabolic diseases such as cancer.