dc.contributorUniversidade Federal de Goiás (UFG)
dc.contributorUniversidade Estadual Paulista (Unesp)
dc.date.accessioned2014-05-27T11:29:30Z
dc.date.available2014-05-27T11:29:30Z
dc.date.created2014-05-27T11:29:30Z
dc.date.issued2013-05-16
dc.identifierBMC Microbiology, v. 13, n. 1, 2013.
dc.identifier1471-2180
dc.identifierhttp://hdl.handle.net/11449/75406
dc.identifier10.1186/1471-2180-13-107
dc.identifierWOS:000319513000001
dc.identifier2-s2.0-84877614413
dc.identifier2-s2.0-84877614413.pdf
dc.identifier0000-0002-8059-0826
dc.description.abstractBackground: 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. © 2013 de Oliveira et al.; licensee BioMed Central Ltd.
dc.languageeng
dc.relationBMC Microbiology
dc.relation2.829
dc.relation1,242
dc.rightsAcesso aberto
dc.sourceScopus
dc.subjectMalate synthase
dc.subjectParacoccidioides spp
dc.subjectProtein-protein interactions
dc.subjectmalate synthase
dc.subjectamino acid analysis
dc.subjectamino acid sequence
dc.subjectantibody production
dc.subjectantibody specificity
dc.subjectcomputer model
dc.subjectcomputer program
dc.subjectconfocal microscopy
dc.subjectcontrolled study
dc.subjectFar Western blotting
dc.subjectgenetic transcription
dc.subjectheterologous expression
dc.subjectimmunofluorescence
dc.subjectin vivo study
dc.subjectligand binding
dc.subjectlung alveolus cell
dc.subjectmacrophage
dc.subjectmass spectrometry
dc.subjectmolecular docking
dc.subjectmolecular dynamics
dc.subjectmolecular interaction
dc.subjectmolecular model
dc.subjectmycelium
dc.subjectParacoccidioides
dc.subjectprotein analysis
dc.subjectprotein binding
dc.subjectprotein determination
dc.subjectprotein function
dc.subjectprotein interaction
dc.subjectprotein protein interaction
dc.subjectprotein purification
dc.subjectprotein secretion
dc.subjectSaccharomyces cerevisiae
dc.subjectsequence homology
dc.subjectsignal transduction
dc.subjecttwo hybrid system
dc.subjectyeast
dc.subjectFungi
dc.subjectMurinae
dc.titleIntermolecular interactions of the malate synthase of Paracoccidioides spp
dc.typeArtículos de revistas


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