Proteomic profile response of Paracoccidioides lutzii to the antifungal argentilactone

Abstract

The dimorphic fungi Paracoccidioides spp. are the etiological agents of paracoccidioidomycosis (PCM), a mycosis of high incidence in Brazil. The toxicity of drug treatment and the emergence of resistant organisms have led to research for new candidates for drugs. In this study, we demonstrate that the natural product argentilactone was not cytotoxic or genotoxic to MRC5 cells at the IC50 concentration to the fungus. We also verified the proteomic profile of Paracoccidioides lutzii after incubation with argentilactone using a label free quantitative proteome nanoUPLC-MSE. The results of this study indicated that the fungus has a global metabolic adaptation in the presence of argentilactone. Enzymes of important pathways, such as glycolysis, the Krebs cycle and the glyoxylate cycle, were repressed, which drove the metabolism to the methylcytrate cycle and beta-oxidation. Proteins involved in cell rescue, defense and stress response were induced. In this study, alternative metabolic pathways adopted by the fungi were elucidated, helping to elucidate the course of action of the compound studied.

Argentilactone Molecular Targets in Paracoccidioides brasiliensis Identified by Chemoproteomics

“…Argentilactone, the major component of H. ovalifolia essential oil, was able to inhibit the growth of P. lutzii without promoting cytotoxic and genotoxic effects on MRC5 cells (14). Here, we expanded the evaluation of MIC and minimum fungicidal concentration (MFC) for other species, such as P. brasiliensis, P. americana, and P. restrepiensis.…”

“…The elucidation of the molecular target is an essential step for developing molecules with bioactive potentials (17). The proteomic and transcriptional responses of P. lutzii to argentilactone were previously evaluated by Prado et al (14) and Araújo et al (16). Although these analyses provided important information, the interactional level of argentilactone and proteins in P. brasiliensis is unknown.…”

“…These proteins may have interacted directly or indirectly, resulting in a total of 109 proteins identified. Among the argentilactone-interacting proteins, 12 of them were downregulated and 6 were upregulated in response to argentilactone, according to the proteomics analysis of P. lutzii (14). Concerning the transcript levels, 8 genes were upregulated in the presence of argentilactone (16) (see Table S1 in the supplemental material).…”

“…In addition, aiming to evaluate if argentilactone induces DNA damage in human cells, the comet assay was performed for MRC5 cells treated with different concentrations of argentilactone. In the MRC5 cells, argentilactone did not induce DNA damage, suggesting that this compound is safe to humans (14).…”

Manuscript title: antifungal proteins from moulds: analytical tools and potential application to dry-ripened foods

“…Advances in mass spectrometry (MS)-based proteomic methods have enabled in-depth analysis of the protein levels altered in response to an antifungal challenge. Label-free methods have been utilized to profile the AFPs response within both resistant and sensitive fungi, enabling the identification of resistance mechanisms as well as susceptible targets (Delgado et al 2015b;Prado et al 2015;Delgado et al 2016).…”