Abstract

The aim of this study is to select a cisplatin-resistant Saccharomyces cerevisiae strain to look for new molecular markers of resistance and the identification of mechanisms/interactions involved. A resistant strain was obtained after 80 days of cisplatin exposure. Then, total protein extraction, purification, and identification were carried out, in wild-type (wt) and resistant strains, by tandem mass spectrometry using a “nano HPLC-ESI-MS/MS” ion trap system. The increase in the exponentially modified protein abundance index (emPAI) (resistant vs wt strains) was calculated to study the increase in protein expression. “Genemania” software (http://www.Genemania.org/) was used to compare the effects, functions, and protein interactions. KEGG tool was used for metabolic pathway analysis. Data are available via ProteomeXchange with identifier PXD020665. The cisplatin-resistant strain showed 2.5 times more resistance than the wt strain for the inhibitory dose 50% (ID50) value (224 μg/ml vs 89.68 μg/ml) and 2.78 times more resistant for the inhibitory dose 90% (ID90) value (735.2 μg/ml vs 264.04 μg/ml). Multiple deregulated proteins were found in the glutathione and carbon metabolism, oxidative phosphorylation, proteasome, glycolysis and gluconeogenesis, glyoxylate metabolism, fatty acid degradation pathway, citric acid cycle, and ribosome. The most overexpressed proteins in the cisplatin-resistant strain were related to growth and metabolism (QCR2, QCR1, ALDH4, ATPB, ATPA, ATPG, and PCKA), cell structure (SCW10), and thermal shock (HSP26). The results suggest that these proteins could be involved in cisplatin resistance. The resistance acquisition process is complex and involves the activation of multiple mechanisms that interact together.

Key points

• Identification of new proteins/genes related to cisplatin resistance

• Increased expression of QCR2/QCR1/ALDH4/ATPB/ATPA/SCW10/HSP26/ATPG and PCKA proteins

• Multiple molecular mechanisms that interact together are involved in resistance

Graphical abstract

中文翻译：

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酿酒酵母中与顺铂耐药相关的新蛋白的鉴定

摘要

这项研究的目的是选择一种耐顺铂啤酒酵母菌株以寻找新的抗性分子标记并鉴定涉及的机制/相互作用。顺铂暴露80天后获得了耐药菌株。然后，使用“纳米HPLC-ESI-MS / MS”离子阱系统通过串联质谱在野生型（wt）和抗性菌株中进行总蛋白的提取，纯化和鉴定。计算出指数修饰的蛋白质丰度指数（emPAI）的增加（抗性与wt菌株），以研究蛋白质表达的增加。使用“ Genemania”软件（http://www.Genemania.org/）比较效果，功能和蛋白质相互作用。KEGG工具用于代谢途径分析。数据可通过ProteomeXchange获得，其标识符为PXD020665。顺铂耐药菌株显示2。抑制剂量50％（ID50）值（224μg/ ml对89.68μg/ ml）的抗性比wt菌株高5倍，对抑制剂量90％（ID90）值（735.2μg/ ml对抑制力）的2.78倍264.04μg/ ml）。在谷胱甘肽和碳代谢，氧化磷酸化，蛋白酶体，糖酵解和糖异生，乙醛酸代谢，脂肪酸降解途径，柠檬酸循环和核糖体中发现了多种失调的蛋白质。顺铂耐药菌株中最过度表达的蛋白质与生长和代谢（QCR2，QCR1，ALDH4，ATPB，ATPA，ATPG和PCKA），细胞结构（SCW10）和热休克（HSP26）有关。结果表明这些蛋白可能与顺铂耐药有关。

关键点

•鉴定与顺铂耐药性有关的新蛋白质/基因

•QCR2 / QCR1 / ALDH4 / ATPB / ATPA / SCW10 / HSP26 / ATPG和PCKA蛋白的表达增加

•抵抗力涉及多种相互作用的分子机制

图形概要