br Experimental design materials and methods br Acknowledgme

Experimental design, materials and methods
Acknowledgments This work was supported by a Grant from the Swedish Cancer Society.
Specifications Table
Value of the data
Data, experimental design, materials and methods Using a quantitative shotgun proteomic approach (nLC–ESI–MS/MS) we have previously identified in these dataset YO-01027 with potential role in the regulation of virulence in the pathogenic fungus Paracoccidioides brasiliensis[1]. The experimental workflow was designed to compare attenuated and virulent Pb18 strains in MM or RM media, to identify potential proteins involved in the virulence regulation. Each strain was grown in biological duplicates, proteins were extracted, digested with trypsin, labeled with light and heavy formaldehyde, and each biological replicate was analyzed by LC–MS/MS in technical triplicates. Raw data were processed and analyzed in PEAKS Studio 7.0 (Bioinformatics Solutions Inc.) and proteins were quantified by the ratios of heavy/light peptide intensities. qPCR assays were performed to validate the LC–MS/MS data (Fig. 1).
Acknowledgments The authors acknowledge the financial support from FAPESP (Fundação de Amparo à Pesquisa do Estado de São Paulo/Brazil) Proc. 2011/14392-2, 2014/13961-1 to WLB and Proc. 2012/19321-9 to AKT. The financial support from CNPq (Conselho Nacional de Desenvolvimento Científico e Tecnologico/Brazil) Proc. 478023/2013-8 to WLB is also acknowledged.
Data EdAG has been shown to irreversibly glutathionylate and inhibit Grx\'s which play a critical role in the glutathionylation and deglutathionylation of many proteins. Grx\'s are important for many cellular regulatory processes [1]. Many other redoxins that contain active site cys residues in GSH binding sites, or other proteins with nucleophilic cys residues, may be targets for EdAG as well. Collectively, these reactions could contribute to the clearance, distribution, or toxicity of busulfan. Further studies on the mechanism of busulfan and its metabolite EdAG are required. However, EdAG is not commercially available and requires synthesis from GSH. The NMR characterization reported here will facilitate future efforts to synthesize EdAG. In addition, Reference [1] documents the relative specificity of EdAG for cys residues in GSH binding sites, and the mass spectral data included here demonstrate the lack of reaction of EdAG at other cys residues and they demonstrate the apparent oxidation of Grx\'s, independent from EdAG treatment. A scheme depicting the overall two step synthesis is shown in the Figs. 1 and 2 shows the homo-and heteronuclear correlations characterized by NMR. Figs. 3–7 are 2D-homo- and heteronuclear correlation spectra as indicated (Figs. 8–12).
Acknowledgements This work was supported by the Department of Medicinal chemistry, University of Washington (Grant No. R01A182963).
Data, experimental design, materials and methods
Acknowledgments This work was supported by a Grant-in-Aid from the National High Technology Research and Development Program (2012AA02A708), the National Natural Science Foundation of China (31070028 and 31270149), and the National Basic Research Program of China (2011CBA00801). This work also received technical support from the Core Facility Center for Life Sciences, University of Science and Technology of China. The authors do not have any possible conflicts of interest.
Specifications table
Value of the data
Data, experimental design, materials and methods
Materials and methods
Acknowledgments This work was funded by the QUT (Queensland University of Technology) Higher Degree Research Support and a QUT ECARD grant awarded to Peter Prentis. We express our thanks to all the group members of Physiological Genomics Lab at QUT for their valuable guidance and support.
Data
Experimental design and materials