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      Proteomic Profile of Cryptococcus neoformans Biofilm Reveals Changes in Metabolic Processes

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          Abstract

          Cryptococcus neoformans, a pathogenic yeast, causes meningoencephalitis, especially in immunocompromised patients, leading in some cases to death. Microbes in biofilms can cause persistent infections, which are harder to treat. Cryptococcal biofilms are becoming common due to the growing use of brain valves and other medical devices. Using shotgun proteomics we determine the differences in protein abundance between biofilm and planktonic cells. Applying bioinformatic tools, we also evaluated the metabolic pathways involved in biofilm maintenance and protein interactions. Our proteomic data suggest general changes in metabolism, protein turnover, and global stress responses. Biofilm cells show an increase in proteins related to oxidation–reduction, proteolysis, and response to stress and a reduction in proteins related to metabolic process, transport, and translation. An increase in pyruvate-utilizing enzymes was detected, suggesting a shift from the TCA cycle to fermentation-derived energy acquisition. Additionally, we assign putative roles to 33 proteins previously categorized as hypothetical. Many changes in metabolic enzymes were identified in studies of bacterial biofilm, potentially revealing a conserved strategy in biofilm lifestyle.

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          Most cited references61

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          Gene Ontology: tool for the unification of biology

          Genomic sequencing has made it clear that a large fraction of the genes specifying the core biological functions are shared by all eukaryotes. Knowledge of the biological role of such shared proteins in one organism can often be transferred to other organisms. The goal of the Gene Ontology Consortium is to produce a dynamic, controlled vocabulary that can be applied to all eukaryotes even as knowledge of gene and protein roles in cells is accumulating and changing. To this end, three independent ontologies accessible on the World-Wide Web (http://www.geneontology.org) are being constructed: biological process, molecular function and cellular component.
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            Large-scale analysis of the yeast proteome by multidimensional protein identification technology.

            We describe a largely unbiased method for rapid and large-scale proteome analysis by multidimensional liquid chromatography, tandem mass spectrometry, and database searching by the SEQUEST algorithm, named multidimensional protein identification technology (MudPIT). MudPIT was applied to the proteome of the Saccharomyces cerevisiae strain BJ5460 grown to mid-log phase and yielded the largest proteome analysis to date. A total of 1,484 proteins were detected and identified. Categorization of these hits demonstrated the ability of this technology to detect and identify proteins rarely seen in proteome analysis, including low-abundance proteins like transcription factors and protein kinases. Furthermore, we identified 131 proteins with three or more predicted transmembrane domains, which allowed us to map the soluble domains of many of the integral membrane proteins. MudPIT is useful for proteome analysis and may be specifically applied to integral membrane proteins to obtain detailed biochemical information on this unwieldy class of proteins.
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              DTASelect and Contrast: tools for assembling and comparing protein identifications from shotgun proteomics.

              The components of complex peptide mixtures can be separated by liquid chromatography, fragmented by tandem mass spectrometry, and identified by the SEQUEST algorithm. Inferring a mixture's source proteins requires that the identified peptides be reassociated. This process becomes more challenging as the number of peptides increases. DTASelect, a new software package, assembles SEQUEST identifications and highlights the most significant matches. The accompanying Contrast tool compares DTASelect results from multiple experiments. The two programs improve the speed and precision of proteomic data analysis.
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                Author and article information

                Journal
                J Proteome Res
                J. Proteome Res
                pr
                jprobs
                Journal of Proteome Research
                American Chemical Society
                1535-3893
                1535-3907
                19 January 2015
                19 January 2014
                07 March 2014
                : 13
                : 3
                : 1545-1559
                Affiliations
                []Department of Chemical Physiology, The Scripps Research Institute , North Torrey Pines Road, Suite 11, La Jolla, California 92037, United States
                []Universidade Federal do Rio Grande do Sul , Avenida Bento Gonçalves, 9500 prédio 43431 sala 214, Porto Alegre, RS 91501-970, Brasil
                Author notes
                Article
                10.1021/pr401075f
                3993910
                24467693
                06909b41-2d5c-4d53-b715-7c7e142baac5
                Copyright © 2014 American Chemical Society
                History
                : 29 October 2013
                Funding
                National Institutes of Health, United States
                Categories
                Article
                Custom metadata
                pr401075f
                pr-2013-01075f

                Molecular biology
                cryptococcus neoformans,biofilm,shotgun proteomics,metabolic changes,resistance
                Molecular biology
                cryptococcus neoformans, biofilm, shotgun proteomics, metabolic changes, resistance

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