The adhesin potential of PbMLS was demonstrated through Far-Western blot, ELISA and binding assays. These showed that the recombinant protein recognized the ECM proteins, fibronectin and SCH 900776 supplier types I and IV collagen, as well as pulmonary epithelial cells. This event indicates that PbMLS can play a role in the interaction of the fungus with host components. Studies have reported the capaCity of P. brasiliensis for adhesion and invasion [9, 15]. This is the first glyoxylate cycle enzyme identified on the fungal surface and released extracellularly which possesses the ability

to bind to ECM proteins. The definition of PbMLSr as a surface-exposed ECM-binding protein, with an unknown mechanism for secretion from the cell or sorting proteins to cellular membrane, suggests that PbMLSr is compatible with

anchorless adhesions [36, 20]. In these types of adhesions, proteins are reassociated on the cellular surface after being secreted to execute their biological functions [36]. The presence of PbMLS in the culture filtrate harvested after 24 and 36 h, and 7 and 14 days of growth selleck chemicals confirmed that it is truly a secreted protein. The presence of PbMLS in SDS-extracted cell-wall protein fraction indicates that PbMLS is associated with the cell surface through weak interactions. Taken together these results provide evidence that PbMLS may be transported out of the cell through the cell wall to be localized on the outer surface of the cell. Reports have described the

presence of some enzymes of the glycolytic pathway on the cell surface in P. brasiliensis as well as in other pathogens [16–19, 37, 38]. The presence of these housekeeping enzymes in unusual locations often correlates with their learn more ability to perform alternative functions such as adherence/invasion of the host cells [38, 18]. The ability of anti-adhesin antibodies to confer protection by blocking microbial attachment to host cells is being explored as a vaccination strategy in several microbial diseases [39–43]. The identification of the PbMLS as a probable adhesin has several implications. Understanding the consequences of the binding of PbMLS to host cells will lead to improved understanding of the initial events during infection. Further insights into the role of the PbMLS in the host-pathogen interaction could contribute Clomifene to the design of novel therapeutic strategies for PCM control. Although PCM infection starts by inhalation of airborne propagules of the mycelia phase, as conidia, which reach the lungs and differentiates into the yeast phase [2], we performed experiments just with yeast cells since this is the phase found inside the host. Is important emphasize that Pbmls transcript is also present in the mycelium phase as described [44, 45]. The results of confocal laser scanning microscopy demonstrated differences in the accumulation of PbMLS among P.