XI International Meeting on Paracoccidioidomycosis

XI International Meeting on Paracoccidioidomycosis

Resume:18-2

Poster (Painel)

18-2	*PROTEOMIC ANALYSIS AND FUNCTIONAL CLASSIFICATION OF CELL WALL PROTEINS OF Paracoccidioides brasiliensis* YEAST FORM**
Authors:	Simone Schneider Weber (LBM / UFG - LABORATÓRIO DE BIOLOGIA MOLECULAR) ; Ana Flávia Alves Parente (LBM / UFG - LABORATÓRIO DE BIOLOGIA MOLECULAR) ; Hellen Kupffer Philippsen (LBM / UFG - LABORATÓRIO DE BIOLOGIA MOLECULAR) ; Alexandre Melo Bailão (LBM / UFG - LABORATÓRIO DE BIOLOGIA MOLECULAR) ; Célia Maria de Almeida Soares (LBM / UFG - LABORATÓRIO DE BIOLOGIA MOLECULAR)

Abstract

Paracoccidioides brasiliensis, the causative agent of Paracoccidioidomycosis, can grow as mycelial or yeast. Host-fungal interactions are often mediated via surface-associated proteins and the identification of these molecules is an important goal of fungal proteomics. The cell wall proteins (CWPs) of P. brasiliensis play a key role in morphogenesis and pathogenesis and might be potential targets for new specific antifungical drugs. The cell wall of fungal pathogens is critical for adhesion to host tissues, representing an essential step during infection. We used two-dimensional electrophoresis to screen the cell wall proteins, which are extracted by cell wall fractionation according to the type of interactions that they establish with other structural components. The standardization was based in literature data for the model Saccharomyces cerevisiae and Candida albicans with adaptations. Mass spectra of tryptic peptides were taken with a MALDI-Q-TOF and identified by Mascot software. The identified proteins were described in functional categories according to the MIPS Functional Catalogue Database (FunCatDB). One gel of each fraction (1 and 2) was used as a reference 2-DE map. Our prior results of cell wall proteins extracted with DTT/SDS and NaOH include several proteins of interest that are important to understand the fungal pathogenesis. In this study, were identified by mass spectrometry some proteins previously described in the P. brasiliensis cell wall as enolase, glyceraldehyde-3-phosphate dehydrogenase, woronin body major protein and triosephosphate isomerase. The enolase, GAPDH and TPI have been found, in this fungus, mediate adhesion to extracellular matrix components. We report the identification of some non-classical CWPs, as elongation factors, glycolytic enzymes and heat shock proteins, at cell surface, suggesting the presence of secretory pathways. For instance, the malato dehydrogenase was found in the cell wall of P. brasiliensis and we suppose that could be due secretion process, since MDH was described in peroxissomes, and this protein is predicted to have signal peptide according to SignalP software. Here, we have report the identification of cell wall proteome of P. brasiliensis yeast form.

Keyword: Paracoccidioides brasiliensis, PROTEOME, CELL WALL, MASS SPECTROMETRY

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