Antifungal resistance is an emerging problem and one of the reasons for treatment failure of invasive aspergillosis (IA). Voriconazole has become a standard therapeutic for the treatment of this often fatal infection. We studied the differentially expressed proteins as a response of Aspergillus fumigatus to voriconazole by employing the two-dimensional difference gel electrophoresis (DIGE) technique. Due to addition of drug, a total of 135 differentially synthesized proteins were identified by MALDI-TOF/TOF-mass spectrometry. In particular, the level of proteins involved in the general stress response and cell detoxification increased prominently. In contrast, cell metabolism and energy proteins were down-regulated, which suggests the cellular effort to maintain balance in energy utilization while trying to combat the cellular stress exerted by the drug. We detected several so-far uncharacterized proteins which may play a role in stress response and drug metabolism and which could be future targets for antifungal treatment. A mutant strain, which is deleted in the cross-pathway control gene cpcA, was treated with voriconazole to investigate the contribution of the general control of amino acid biosynthesis to drug resistance. We compared the mutant strain's protein expression profile with the wild-type strain. The absence of CpcA led to an increased resistance to voriconazole and a reduced activation of some general stress response proteins, while the transcript level of the triazole target gene erg11A (cyp51A) remained unchanged. In contrast, the sensitivity of strain ΔcpcA to terbinafine and amphotericin B was slightly increased. These findings imply a role of CpcA in the cellular stress response to azole drugs at the post transcriptional level.

中文翻译：

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烟曲霉对伏立康唑抗真菌药物反应的蛋白质组学分析。

抗真菌药耐药性是一个新出现的问题，是浸润性曲霉病（IA）治疗失败的原因之一。伏立康唑已成为治疗这种致命性感染的标准疗法。我们通过利用二维差异凝胶电泳（DIGE）技术研究了差异表达的蛋白质作为烟曲霉对伏立康唑的反应。由于添加了药物，通过MALDI-TOF / TOF-质谱法鉴定出总共135种差异合成的蛋白质。特别地，参与一般应激反应和细胞排毒的蛋白质水平显着增加。相反，细胞代谢和能量蛋白被下调，这表明细胞努力维持能量利用的平衡，同时努力对抗药物施加的细胞压力。我们检测到了一些迄今未表征的蛋白，这些蛋白可能在应激反应和药物代谢中起作用，并且可能成为抗真菌治疗的未来目标。用伏立康唑处理了在交叉途径控制基因cpcA中缺失的突变菌株，以研究氨基酸生物合成的一般控制对耐药性的贡献。我们将突变菌株的蛋白质表达谱与野生型菌株进行了比较。CpcA的缺乏导致对伏立康唑的耐药性增加，某些一般应激反应蛋白的激活降低，而三唑靶基因erg11A（cyp51A）的转录水平保持不变。相比之下，ΔcpcA菌株对特比萘芬和两性霉素B的敏感性略有提高。这些发现暗示了CpcA在转录后水平上对唑类药物的细胞应激反应中的作用。