Dataset Information

Glycoconjugates and polysaccharides of fungal cell wall and activation of immune system.

ABSTRACT: Glycoproteins, glycosphingolipids and polysaccharides exposed at the most external layers of the wall are involved in several types of interactions of fungal cells with the exocellular environment. These molecules are fundamental building blocks of organisms, contributing to the structure, integrity, cell growth, differentiation and signaling. Several of them are immunologically active compounds with potential as regulators of pathogenesis and the immune response of the host. Some of these structures can be specifically recognized by antibodies from patients' sera, suggesting that they can be also useful in the diagnosis of fungal infections.

SUBMITTER: Pinto MR

PROVIDER: S-EPMC3768395 | biostudies-literature | 2008 Apr

REPOSITORIES: biostudies-literature

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Glycoconjugates and polysaccharides of fungal cell wall and activation of immune system.

Pinto M R MR Barreto-Bergter E E Taborda C P CP

Brazilian journal of microbiology : [publication of the Brazilian Society for Microbiology] 20080401 2

Glycoproteins, glycosphingolipids and polysaccharides exposed at the most external layers of the wall are involved in several types of interactions of fungal cells with the exocellular environment. These molecules are fundamental building blocks of organisms, contributing to the structure, integrity, cell growth, differentiation and signaling. Several of them are immunologically active compounds with potential as regulators of pathogenesis and the immune response of the host. Some of these struc ...[more]

PMID: 24031202

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Project description:This study was a proteomic and secretomic survey of the Gram-negative saprophytic bacterium Cellvibio japonicus during growth using fungal polysaccharides or authentic fungal biomass as a sole nutrient source. The purified polysaccharides were beta-glucan (derived from yeast) and alpha chitin (derived from shrimp shells). The authentic fungal biomass came from Saccharomyces cerevisiae and Aspergillus nidulans. The proteome and secretome samples were taken during mid-exponential growth in biological triplicate. Bacterial cells were homogenized via probe sonication and reduced and alkylated followed by trypsin digestion using a suspension trap (S-trap). Supernatants were reduced and alkylated and followed by trypsin digestion via MStern Blotting. An SPQC was made by combining equal volumes of each sample for both sample types. After lyophilization and reconstitution, 2 ul of cell digests and 15 ul of supernatant digests were loaded onto Evotips. Quantitative LC/MS/MS was performed using an Evosep One LC coupled to a Thermo Orbitrap Astral via a Nanospray Flex ionization source. Samples were block randomized and interspersed with the SPQC replicates. The Evosep used a 60 sample-per-day method with a Pepsep 8 cm x 150 um column (1.5 um particle size) with a PepSep Sprayer and stainless steel (30 um) emitter. The MS analysis used a 240,000 resolution precursor ion (MS1) scan from 380-980 m/z, AGC target of 500% and maximum injection time (IT) of 50 ms, collected every 0.6 s in centroid mode. MS/MS was performed using a DIA method with default charge state = 3, precursor mass range of 380-980, 10 m/z isolation windows, AGC target of 500% and maximum IT of 20 ms, and a NCE of 28. An RF lens of 40% was used for MS1 and DIA scans. Raw MS data was converted to .htrms format using HTRMS converter and processed in Spectronaut 19 (Biognosys). A spectral library was built using direct-DIA searches of all DIA analyses using a Cellvibrio japonicus database, downloaded from Uniprot on 10/29/2024 and appended contaminant sequences using FragPipeSearch settings included N-terminal trypsin/P specificity up to 2 missed cleavages; peptide length from 7-52 amino acids with the following modifications: acetyl (N-term) and carbamidomethyl(Cys). For DIA analysis, default extraction, calibration, identification, and protein inference settings were used.

Dataset Information

Glycoconjugates and polysaccharides of fungal cell wall and activation of immune system.

Publications

Glycoconjugates and polysaccharides of fungal cell wall and activation of immune system.

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