Characterization and functional analysis of the Î²-1,3-glucanosyltransferase 3 of the human pathogenic fungus<i>Paracoccidioides brasiliensis</i>

Castro, Nadya da Silva; Castro, Kelly Pacheco de; Orlandi, Ivan; Feitosa, Luciano dos Santos; Kmetzsch, Lívia; Vainstein, Marilene Henning; Báo, Sônia Nair; Vai, Marina; Soares, Célia Maria de Almeida

doi:10.1111/j.1567-1364.2008.00463.x

Cited by 20 publications

(11 citation statements)

References 40 publications

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“…For the ultrastructural and immunocytochemistry studies, previously described protocols were employed [76,81,82]. The yeast cells were fixed in solution containing 2% (v/v) glutaraldehyde, 2% (w/v) paraformaldehyde, and 3% (w/v) sucrose in 0.1 M sodium cacodylate buffer pH 7.2.…”

Section: Methodsmentioning

confidence: 99%

Analysis of Paracoccidioides secreted proteins reveals fructose 1,6-bisphosphate aldolase as a plasminogen-binding protein

et al. 2015

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BackgroundDespite being important thermal dimorphic fungi causing Paracoccidioidomycosis, the pathogenic mechanisms that underlie the genus Paracoccidioides remain largely unknown. Microbial pathogens express molecules that can interact with human plasminogen, a protein from blood plasma, which presents fibrinolytic activity when activated into plasmin. Additionally, plasmin exhibits the ability of degrading extracellular matrix components, favoring the pathogen spread to deeper tissues. Previous work from our group demonstrated that Paracoccidioides presents enolase, as a protein able to bind and activate plasminogen, increasing the fibrinolytic activity of the pathogen, and the potential for adhesion and invasion of the fungus to host cells. By using proteomic analysis, we aimed to identify other proteins of Paracoccidioides with the ability of binding to plasminogen.ResultsIn the present study, we employed proteomic analysis of the secretome, in order to identify plasminogen-binding proteins of Paracoccidioides, Pb01. Fifteen proteins were present in the fungal secretome, presenting the ability to bind to plasminogen. Those proteins are probable targets of the fungus interaction with the host; thus, they could contribute to the invasiveness of the fungus. For validation tests, we selected the protein fructose 1,6-bisphosphate aldolase (FBA), described in other pathogens as a plasminogen-binding protein. The protein FBA at the fungus surface and the recombinant FBA (rFBA) bound human plasminogen and promoted its conversion to plasmin, potentially increasing the fibrinolytic capacity of the fungus, as demonstrated in fibrin degradation assays. The addition of rFBA or anti-rFBA antibodies was capable of reducing the interaction between macrophages and Paracoccidioides, possibly by blocking the binding sites for FBA. These data reveal the possible participation of the FBA in the processes of cell adhesion and tissue invasion/dissemination of Paracoccidioides.ConclusionsThese data indicate that Paracoccidioides is a pathogen that has several plasminogen-binding proteins that likely play important roles in pathogen-host interaction. In this context, FBA is a protein that might be involved somehow in the processes of invasion and spread of the fungus during infection.

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Section: Methodsmentioning

confidence: 99%

Analysis of Paracoccidioides secreted proteins reveals fructose 1,6-bisphosphate aldolase as a plasminogen-binding protein

et al. 2015

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“…A screening in the Pb01 transcriptome for GPI-anchored sequences (Castro et al, 2005) yielded six expression sequencing tags (ESTs) from possible cell wall structural proteins. A putative GPI-anchored membrane/cell wall β-1,3-glucan elongase capable of mediating fungal cell wall integrity (PbGel3p) was immunolocalized at the yeast surface (Castro et al, 2009). …”

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confidence: 99%

The Paracoccidioides Cell Wall: Past and Present Layers Toward Understanding Interaction with the Host

Puccia¹,

Vallejo²,

Matsuo³

et al. 2011

Front. Microbio.

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The cell wall of pathogenic fungi plays import roles in the interaction with the host, so that its composition and structure may determine the course of infection. Here we present an overview of the current and past knowledge on the cell wall constituents of Paracoccidioides brasiliensis and P. lutzii. These are temperature-dependent dimorphic fungi that cause paracoccidioidomycosis, a systemic granulomatous, and debilitating disease. Focus is given on cell wall carbohydrate and protein contents, their immune-stimulatory features, adhesion properties, drug target characteristics, and morphological phase specificity. We offer a journey toward the future understanding of the dynamic nature of the cell wall and of the changes that may occur when the fungus infects the human host.

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“…Certain P. brasiliensis glycolytic enzymes, such as N -acetyl-β-D-glucosaminidase, α- and β-glucanase, α-amylase, α-glucan synthase, and β-glucanosyltransferase play important functions with respect to the structure of the cell wall. They are also required for the synthesis, and the remodeling of cell walls in response to environmental changes [ 8 - 1 ]. N -glycans are involved in protein folding, intracellular transport, and protection from proteolytic degradation [ 12 ].…”

Section: Introductionmentioning

confidence: 99%

Influence of N-glycans on Expression of Cell Wall Remodeling Related Genes in Paracoccidioides brasiliensis Yeast Cells

Almeida

Antoniêto

Pessoni

et al. 2016

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Paracoccidioidomycosis is the most prevalent systemic mycosis in Latin America. It is caused by the temperature-dependent dimorphic fungus Paracoccidioides brasiliensis. The P. brasiliensis cell wall is a dynamic outer structure, composed of a network of glycoproteins and polysaccharides, such as chitin, glucan and N-glycosylated proteins. These glycoproteins can interact with the host to affect infection rates, and are known to perform other functions. We inhibited N-linked glycosylation using tunicamycin (TM), and then evaluated the expression of P. brasiliensis genes related to cell wall remodeling. Our results suggest that cell wall synthesis related genes, such as β-1,3-glucanosyltransferase (PbGEL3), 1,3-β-D-glucan synthase (PbFKS1), and α-1,4-amylase (PbAMY), as well as cell wall degrading related genes, such as N-acetyl-β-D-glucosaminidase (PbNAG1), α-1,3-glucanase (PbAGN), and β-1,3-glucanase (PbBGN1 and PbBGN2), have their expression increased by the N-glycosylation inhibition, as detected by qRT-PCR. The observed increases in gene expression levels reveal possible compensatory mechanisms for diminished enzyme activity due to the lack of glycosylation caused by TM.

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Characterization and functional analysis of the Î²-1,3-glucanosyltransferase 3 of the human pathogenic fungusParacoccidioides brasiliensis

Cited by 20 publications

References 40 publications

Analysis of Paracoccidioides secreted proteins reveals fructose 1,6-bisphosphate aldolase as a plasminogen-binding protein

Analysis of Paracoccidioides secreted proteins reveals fructose 1,6-bisphosphate aldolase as a plasminogen-binding protein

The Paracoccidioides Cell Wall: Past and Present Layers Toward Understanding Interaction with the Host

Influence of N-glycans on Expression of Cell Wall Remodeling Related Genes in Paracoccidioides brasiliensis Yeast Cells

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