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首页> 外文期刊>Renewable Agriculture and Food Systems >Label-free protein quantification of sodium butyrate treated CHO cells by ESI-UHR-TOF-MS
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Label-free protein quantification of sodium butyrate treated CHO cells by ESI-UHR-TOF-MS

机译:通过ESI-UHR-TOF-MS对丁酸钠处理CHO细胞的无标记蛋白质定量

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摘要

Effects of butyrate on CHO producer cells are contradictory, promoting productivity and at the same time repressing proliferation. Though in previous omics studies the background of butyrate impact on producer cells has been investigated, the knowledge about the mechanism is still very limited. As previous proteomic results on this field are mainly based on 2DE-gels, we conducted a label-free MS quantification, based on fast high resolution ESI-MS and a straight forward software solution, to gain insight in shifted cellular processes of CHO cells 25 h after butyrate treatment. 118 proteins or subunits with significantly altered abundances were identified suggesting changes in carbohydrate, protein metabolic and cell cycle processes. Effects of butyrate on the nucleosome assembly as a known direct epigenetic influence on HDAC activity turned out to be unexpectedly fast and persistent, as confirmed by Western blots of histone-H4 acetylation. Contradictory to increased cell specific productivity, most elements of protein metabolism exhibited decreased levels after butyrate treatment. In comparison to published results some overlap of our label free MS data could be observed but also apparently diverging findings, showing the need for complementary omics techniques for a holistic view on cellular processes such as response to butyrate.
机译:丁酸盐对CHO生产者细胞的影响是矛盾的,促进生产率,同时抑制增殖。虽然在先前的OMICS研究中,研究了对生产者细胞的影响,但已经研究了对机制的了解仍然非常有限。由于此领域的先前蛋白质组学结果主要基于2DE-GEL,我们基于快速高分辨率ESI-MS和直线软件解决方案进行了无标签MS量化,以获得CHO细胞25的移位蜂窝过程中的洞察力丁酸盐处理后。鉴定了118个蛋白质或亚基,具有显着改变的丰富的丰富,表明碳水化合物,蛋白质代谢和细胞周期过程的变化。丁酸盐对核心组装作为已知的直接表观遗传对HDAC活性的影响,原来是出乎意料的快速和持续的,如通过组蛋白-H4乙酰化的蛋白质印迹证实。对细胞比生产力提高的矛盾,蛋白质代谢的大多数元素表现出降低的丁酸处理后水平降低。与公布结果相比,可以观察到我们标签免费MS数据的一些重叠,但也显然发散结果,表明需要对诸如对丁酸丁酸反应的细胞过程进行整体视图的互补常规观察。

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