Resumo

The thermally dimorphic fungus Paracoccidioides spp. is the cause of the most endemic mycosis in Latin America, the Paracoccidioidomycosis (PCM). During infection Paracoccidioides spp. needs a variety of elements to survive with in the host. The metallic micronutrients are essential elements required for the cell homeostasis in virtually all organisms. After genomic era, proteomics have been used as an important approach to study the gene functions in a large scale. The proteomic studies aim to analyze various proteins aspects since functional up to structural ones. The metalloproteomic is study metal-binding proteins. The interaction with metal is essential to proteins structure formation, catalytic activity in all biologic systems. Zinc and copper as essential metal which are present in many metalloproteins. The zinc is one of the most important metal for biological systems playing role in more than 300 enzymes and biochemical processes. Copper is a cofactor of many oxidoreduction enzymes of the respiratory chain. Recent studies have shown that the homeostasis of those metals are important for fungal pathogenesis. This work purpose to identify zinc- and copper-proteins of Paracoccidioides spp. yeast cells. For that, it has been used immobilized metal affinity chromatography (IMAC-Select Affinity Gel) coupled nanoUPLC-MSE. For that IMAC resin was load with zinc or copper. The metalloproteins were load on the metal loaded resins and then eluted in buffers with crescent denaturating strengths. Zinc binding proteins were unbounded from resin at 20 mM of imidazole. On the other hand, copper proteins bounded more tightly to the Cu-IMAC. Most of them were extracted at 200 mM of imidazole and with EDTA wash. The identification of the purified metalloproteins is under progress. The characterization of copper and zinc proteins from Paracoccidioides spp. can bring new insights of metal homeostasis and pathogenesis of this fungus.