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Tipo do documento: Artigo
Título: Intermolecular interactions of the malate synthase of Paracoccidioides spp
Autor: Oliveira, Karine Martins de
Silva Neto, Benedito Rodrigues da
Rocha, Juliana Alves Parente
Silva, Roosevelt Alves da
Quintino, Guilherme Oliveira
Voltan, Aline Raquel
Giannini, Maria José Soares Mendes
Soares, Célia Maria de Almeida
Pereira, Maristela
Abstract: Background: The fungus Paracoccidioides spp is the agent of paracoccidioidomycosis (PCM), a pulmonary mycosis acquired by the inhalation of fungal propagules. Paracoccidioides malate synthase (PbMLS) is important in the infectious process of Paracoccidioides spp because the transcript is up-regulated during the transition from mycelium to yeast and in yeast cells during phagocytosis by murine macrophages. In addition, PbMLS acts as an adhesin in Paracoccidioides spp. The evidence for the multifunctionality of PbMLS indicates that it could interact with other proteins from the fungus and host. The objective of this study was to identify and analyze proteins that possibly bind to PbMLS (PbMLS-interacting proteins) because protein interactions are intrinsic to cell processes, and it might be possible to infer the function of a protein through the identification of its ligands. Results: The search for interactions was performed using an in vivo assay with a two-hybrid library constructed in S. cerevisiae; the transcripts were sequenced and identified. In addition, an in vitro assay using pull-down GST methodology with different protein extracts (yeast, mycelium, yeast-secreted proteins and macrophage) was performed, and the resulting interactions were identified by mass spectrometry (MS). Some of the protein interactions were confirmed by Far-Western blotting using specific antibodies, and the interaction of PbMLS with macrophages was validated by indirect immunofluorescence and confocal microscopy. In silico analysis using molecular modeling, dynamics and docking identified the amino acids that were involved in the interactions between PbMLS and PbMLS-interacting proteins. Finally, the interactions were visualized graphically using Osprey software. Conclusion: These observations indicate that PbMLS interacts with proteins that are in different functional categories, such as cellular transport, protein biosynthesis, modification and degradation of proteins and signal transduction. These data suggest that PbMLS could play different roles in the fungal cell.
Palavras-chave: Paracoccidioides spp
Protein-protein interactions
Malate synthase
País: Gra-bretanha
Unidade acadêmica: Instituto de Ciências Biológicas - ICB (RG)
Citação: OLIVEIRA, Karine Martins de; SILVA Neto, Benedito Rodrigues da; PARENTE, Juliana Alves; SILVA, Roosevelt Alves da; QUINTINO, Guilherme Oliveira; VOLTAN, Aline Raque; MENDES-GIANNINI, Maria José Soares; SOARES, Célia Maria de Almeida; PEREIRA, Maristela. Intermolecular interactions of the malate synthase of Paracoccidioides spp. BMC Microbiology, London, v. 13, n. 107, p. 1-14, 2013.
Tipo de acesso: Acesso Aberto
Identificador do documento: 10.1186/1471-2180-13-107
Identificador do documento: 10.1186/1471-2180-13-107
Data de publicação: 2013
Aparece nas coleções:IPTSP - Artigos publicados em periódicos

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