Przejdź do zawartości
Merck

Proteome of tolerance fine-tuning in the human pathogen black yeast Exophiala dermatitidis.

Journal of proteomics (2015-07-21)
Donatella Tesei, Gorji Marzban, Martina Marchetti-Deschmann, Hakim Tafer, Elsa Arcalis, Katja Sterflinger
ABSTRAKT

The black yeast Exophiala dermatitidis is a worldwide distributed agent of primary and secondary diseases in both immunocompromised and healthy humans, with a high prevalence in human-made environments. Since thermo-tolerance has a crucial role in the fungus persistence in man-dominated habitat and in its pathogenicity, three incubation temperatures (37, 45, 1 °C) and two time spans (1 h, 1 week) were selected to simulate different environmental conditions and to investigate the effect of temperature on the proteome of E. dermatitidis CBS 525.76. Using a novel protocol for protein extraction from black yeasts, 2-D DIGE could be applied for characterization of changes in total protein spot abundance among the experimental conditions. A total of 32 variable proteins were identified by mass spectrometry. Data about protein functions, localization and pathways were also obtained. A typical stress response under non-optimal temperature could not be observed at the proteome level, whereas a reduction of the metabolic activity, mostly concerning processes as the general carbon metabolism, was detected after exposure to cold. These results suggest that a fine protein modulation takes place following temperature treatment and a repertoire of stable protein might be at the base of E. dermatitidis adaptation to altered growth conditions. E. dermatitidis is a pathogenic black yeast causing neurotropic infections, systemic and subcutaneous disease in a wide range of hosts, including humans. The discovery of the fungus high prevalence in man-made habitats, including sauna facilities, drinking water and dishwashers, generated concern and raised questions about the infection route. In the present work - which is the first contribution on E. dermatitidis proteome - the effect of different temperature conditions on the fungus protein pattern have been analyzed by using a gel-based approach and the temperature responsive proteins have been identified. The absence of a typical stress response following the exposure to non-optimal temperature was detected at the proteome level, along with a general reduction of the metabolic activity after exposure to cold. These results suggest that a very fine regulation of the protein expression as well as adaptations involving a basic set of stable proteins may be at the base of E. dermatitidis enormous ecological plasticity, which plays a role in the fungus distribution, also enabling the transition from natural to human habitat and to the human host.

MATERIAŁY
Numer produktu
Marka
Opis produktu

Sigma-Aldrich
Acetone, ACS reagent, ≥99.5%
Sigma-Aldrich
Acetone, histological grade, ≥99.5%
Sigma-Aldrich
Acetone, natural, ≥97%
Sigma-Aldrich
Acetone, ≥99%, FCC, FG
Sigma-Aldrich
Thiourea, ReagentPlus®, ≥99.0%
Sigma-Aldrich
Urea solution, 40 % (w/v) in H2O
Sigma-Aldrich
Thiourea, ACS reagent, ≥99.0%
Sigma-Aldrich
DL-Dithiothreitol solution, BioUltra, Molecular Biology, ~1 M in H2O
Sigma-Aldrich
D.E.R. 332, used as embedding medium
Sigma-Aldrich
Urea-12C, 99.9 atom % 12C
Sigma-Aldrich
3-[(3-Cholamidopropyl)dimethylammonio]-1-propanesulfonate hydrate, 98%
Sigma-Aldrich
Urea solution, BioUltra, ~8 M in H2O
Sigma-Aldrich
Acetone, ACS reagent, ≥99.5%
Supelco
DL-Dithiothreitol solution, 1 M in H2O
Supelco
Phenol solution, 5000 μg/mL in methanol, certified reference material
Sigma-Aldrich
Bromophenol Blue, ACS reagent
Sigma-Aldrich
Formic acid, ≥95%, FCC, FG
Supelco
Phenol solution, certified reference material, 500 μg/mL in methanol
Sigma-Aldrich
Urea, meets USP testing specifications
Sigma-Aldrich
Urea, suitable for electrophoresis
Sigma-Aldrich
Urea, BioXtra, pH 7.5-9.5 (20 °C, 5 M in H2O)
Sigma-Aldrich
Urea, powder, BioReagent, Molecular Biology, suitable for cell culture
Sigma-Aldrich
Phenol solution, Saturated with 0.01 M citrate buffer, pH 4.3 ± 0.2, BioReagent, Molecular Biology
Sigma-Aldrich
Bromophenol Blue, titration: suitable
Sigma-Aldrich
Phenol, BioXtra, ≥99.5% (GC)
Sigma-Aldrich
Phenol, Molecular Biology
Sigma-Aldrich
Urea, ACS reagent, 99.0-100.5%
Sigma-Aldrich
Liquified Phenol, ≥89.0%
Sigma-Aldrich
Phenol solution, Equilibrated with 10 mM Tris HCl, pH 8.0, 1 mM EDTA, BioReagent, Molecular Biology
Sigma-Aldrich
Phenol, ≥99%