Some PbMLS-interacting proteins were selected for in silico interaction analysis. Proteins were chosen from metabolic pathways such as the glycolytic pathway, the tricarboxylic acid cycle, the methyl citrate cycle and the Seliciclib in vitro glyoxylate cycle because PbMLS participates in the glyoxylate cycle, and the interaction between proteins from different metabolic pathways would be expected. Global energy values for each complex studied showed that there is good complementarity between PbMLS and most PbMLS-interacting proteins. For example, the complexes that involve PbMLS and the proteins

glyceraldehyde-3-phosphate isomerase, malate dehydrogenase, 2-methylcitrate dehydratase and triosephosphate isomerase have global energies that are less than −55 kcal/mol. The global energy values found here were very good. For example, in a recent study of the interactions between D-phosphoglycerate dehydrogenase and phosphoserine aminotransferase from the enteric human parasite Entamoeba histolytica[45], the best global energies were approximately −75 kcal/mol.

Here, the best values were found for fructose 1,6 bisphosphate aldolase and ubiquitin (less than −100 kcal/mol). S. cerevisiae MLS-interacting proteins have already been described. Here, in silico analysis using the S. cerevisiae database showed that PbMLS interacts with other new proteins. The only protein that Vadimezan solubility dmso they share is ubiquitin. This fact and the fact that the interaction between ubiquitin and PbMLS is very stable suggest that this interaction is very important. Ubiquitin is responsible for the conjugation of proteins, marking them for selective degradation via the ubiquitin-proteasome system 26S, a process that is essential in the response to AZD5582 nmr cellular stress. These proteins, however, act through ubiquitination, changing the function, the location and/or the traffic protein, or are targeted for destruction by the 26S proteasome [46]. In conclusion, the molecular interactions that involve proteins located in subcellular compartments facilitate the understanding

of mechanisms that are associated with each interaction. However, proteins are not always at the same location ADAMTS5 in the cell and do not have unique roles [47]. Here, several new PbMLS-interacting proteins from various functional categories were identified, which suggests that their function is diversified beyond the glyoxylate cycle. Conclusions The results of this study indicated that PbMLS interacts with proteins of different functional categories, such as cellular transport, protein biosynthesis, modification and degradation and signal transduction. These data suggest that PbMLS is found in many locations and plays different roles in the fungal cell. Methods Paracoccidioides isolate and growth conditions The fungus Paracoccidioides isolate Pb01 (ATCC MYA-826) was grown, as previously described [39]. The yeast and mycelium phase were grown at 36 and 22 °C, respectively, in Fava–Netto’s medium (1% w/v peptone, 0.