Background

Sporotrichosis is a fungal infection caused by the Sporothrix schenckii complex. In order to colonize the host, the pathogen must neutralize the reactive oxygen species produced by the phagocytic cells during the respiratory burst. Little is known about these mechanisms in S. schenckii.

Aims

To identify the proteins differentially expressed after the exposure of S. schenckii sensu stricto to different concentrations of H₂O₂.

Methods

Yeast cells of S. schenckii sensu stricto were exposed to increasing concentrations of H₂O_2. Proteins differentially expressed in response to oxidative stress were analyzed using two-dimensional polyacrylamide gel electrophoresis (2D-PAGE) and identified by MALDI-MS/MS. RT-PCR assays were performed to evaluate the transcription of genes of the identified proteins.

Results

Concentrations of H₂O₂ as high as 800 mM allowed cell growth, and 200 mM and 400 mM were selected for comparative analysis by 2D-PAGE. This analysis revealed at least five differentially expressed proteins, which were identified as heat shock 70 kDa protein (Hsp70), chaperonin GroEL, elongation factor 1-β (EF1-β), a hypothetical protein, and mitochondrial peroxiredoxin (Prx1). RT-PCR revealed that the transcription of the genes coding for some of these proteins are differentially regulated.

Conclusions

Based on these results, it is proposed that these proteins may be involved in the resistance of S. schenckii to oxidative stress, and play an important role in the fungus survival in the host.

中文翻译：

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响应于过氧化氢诱导的氧化应激，对严格孢子丝菌中的蛋白质进行鉴定。

背景

Sporotrichosis是由Sporothrix schenckii复合体引起的真菌感染。为了定居宿主，病原体必须中和呼吸爆发期间吞噬细胞产生的活性氧。关于S. schenckii中的这些机制知之甚少。

目的

鉴定S. schenckii sensu stricto暴露于不同浓度的H ₂ O ₂后差异表达的蛋白质。

方法

S. chenchenii sensu stricto的酵母细胞暴露于浓度不断增加的H ₂ O _2中。使用二维聚丙烯酰胺凝胶电泳（2D-PAGE）分析响应氧化应激而差异表达的蛋白质，并通过MALDI-MS / MS进行鉴定。进行RT-PCR测定以评估所鉴定蛋白质的基因的转录。

结果

高达800 mM的H ₂ O ₂浓度允许细胞生长，并选择200 mM和400 mM进行2D-PAGE比较分析。该分析揭示了至少五个差异表达的蛋白，这些蛋白被鉴定为热休克70 kDa蛋白（Hsp70），伴侣蛋白GroEL，延伸因子1-β（EF1-β），一种假定蛋白和线粒体过氧化物酶（Prx1）。RT-PCR显示，编码其中某些蛋白质的基因的转录受到差异调节。

结论

基于这些结果，提出这些蛋白质可能参与了德国链球菌对氧化应激的抗性，并且在宿主中的真菌存活中起重要作用。