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Intermolecular interactions of the malate synthase of Paracoccidioides spp

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-regulate...

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Autores principales: de Oliveira, Karine Martins, da Silva Neto, Benedito Rodrigues, Parente, Juliana Alves, da Silva, Roosevelt Alves, Quintino, Guilherme Oliveira, Voltan, Aline Raquel, Mendes-Giannini, Maria José Soares, de Almeida Soares, Célia Maria, Pereira, Maristela
Formato: Online Artículo Texto
Lenguaje:English
Publicado: BioMed Central 2013
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3771410/
https://www.ncbi.nlm.nih.gov/pubmed/23672539
http://dx.doi.org/10.1186/1471-2180-13-107
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author de Oliveira, Karine Martins
da Silva Neto, Benedito Rodrigues
Parente, Juliana Alves
da Silva, Roosevelt Alves
Quintino, Guilherme Oliveira
Voltan, Aline Raquel
Mendes-Giannini, Maria José Soares
de Almeida Soares, Célia Maria
Pereira, Maristela
author_facet de Oliveira, Karine Martins
da Silva Neto, Benedito Rodrigues
Parente, Juliana Alves
da Silva, Roosevelt Alves
Quintino, Guilherme Oliveira
Voltan, Aline Raquel
Mendes-Giannini, Maria José Soares
de Almeida Soares, Célia Maria
Pereira, Maristela
author_sort de Oliveira, Karine Martins
collection PubMed
description 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.
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spelling pubmed-37714102013-09-13 Intermolecular interactions of the malate synthase of Paracoccidioides spp de Oliveira, Karine Martins da Silva Neto, Benedito Rodrigues Parente, Juliana Alves da Silva, Roosevelt Alves Quintino, Guilherme Oliveira Voltan, Aline Raquel Mendes-Giannini, Maria José Soares de Almeida Soares, Célia Maria Pereira, Maristela BMC Microbiol Research Article 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. BioMed Central 2013-05-14 /pmc/articles/PMC3771410/ /pubmed/23672539 http://dx.doi.org/10.1186/1471-2180-13-107 Text en Copyright © 2013 de Oliveira et al.; licensee BioMed Central Ltd. http://creativecommons.org/licenses/by/2.0 This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/2.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.
spellingShingle Research Article
de Oliveira, Karine Martins
da Silva Neto, Benedito Rodrigues
Parente, Juliana Alves
da Silva, Roosevelt Alves
Quintino, Guilherme Oliveira
Voltan, Aline Raquel
Mendes-Giannini, Maria José Soares
de Almeida Soares, Célia Maria
Pereira, Maristela
Intermolecular interactions of the malate synthase of Paracoccidioides spp
title Intermolecular interactions of the malate synthase of Paracoccidioides spp
title_full Intermolecular interactions of the malate synthase of Paracoccidioides spp
title_fullStr Intermolecular interactions of the malate synthase of Paracoccidioides spp
title_full_unstemmed Intermolecular interactions of the malate synthase of Paracoccidioides spp
title_short Intermolecular interactions of the malate synthase of Paracoccidioides spp
title_sort intermolecular interactions of the malate synthase of paracoccidioides spp
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3771410/
https://www.ncbi.nlm.nih.gov/pubmed/23672539
http://dx.doi.org/10.1186/1471-2180-13-107
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