Proteome analysis of Saccharomyces cerevisiae under metal stress by two‐dimensional differential gel electrophoresis

Abstract

The defense mechanism by which cells combat metal stress remains poorly understood. By utilizing a newly developed technique - the differential gel electrophoresis (DIGE) - we evaluated the biological alterations of metal stress on Saccharomyces cerevisiae at its translational level. By simultaneously comparing the differential expression profiles of thousands of proteins as results of 15 different metal treatments, we were able to closely examine the response of a large number of proteins within the yeast proteome towards individual metals, as well as the response of the same proteins towards different metals. This, to our knowledge, is the first case which demonstrates the potential of DIGE as a high-throughput tool for large-scale proteome analysis. From our studies, where yeast cells were exhaustively treated with exogenous metals, 20-30% of all proteins detected showed statistically significant changes. According to different effects (up-/downregulation) of protein expression levels observed, we were able to tentatively divide the 15 metals into three groups. By mass spectrometric analysis, more than 50 protein spots were positively identified, both quantitatively and qualitatively. One of the proteins was identified to be Cu/Zn superoxide dismutase (SOD1), and its expression levels as a result of 15 different metal treatments was further examined in greater details. Significant changes in SOD1 expression were observed throughout all 15 DIGE gels.

Crucial Role of Antioxidant Proteins and Hydrolytic Enzymes in Pathogenicity of Penicillium expansum

“…2 and 3). This may be explained by the difference in exposure time of organisms to stresses in the different experiments (17). The overexpression of antioxidant enzymes often occurs in the studies where cells are exposed to stresses for a short time (always less than 12 h), whereas in our experiment hyphae of P. expansum were treated with borate for 48 -72 h. This result is consistent with the work of Chuang et al (21), who observed that the expression levels of antioxidant enzymes were decreased in the Gram-negative microaerophilic bacterium Helicobacter pylori after long term incubation under oxidative stress.…”

A Proteomics Study of Brassinosteroid Response in Arabidopsis

“…The protein identities of spots of interest are subsequently determined by mass spectrometry. 2-D DIGE has been used widely in quantitative proteomics studies in a wide range of biological and disease systems (35)(36)(37)(38)(39)(40)(41)(42). In plants, 2-D DIGE has been used to identify proteins regulated by UV-B light (43), salt and osmotic stresses (44), cold stress (45), fungal elicitors (46), and gibberellins (47).…”

Differentially expressed proteins of pathogenic Penicillium marneffei in yeast and mycelial phases

“…Cell pellets were resuspended in chilled distilled water and recovered by recentrifugation (3000 r.p.m., 15 min, 4 uC). The pellets were then resuspended in lysis buffer [7 M urea, 2 M thiourea, 4 % CHAPS buffer, 30 mM Tris/HCl (pH 6.8)] at room temperature (Hu et al, 2003). Glass beads (0.4-0.6 mm; Sigma) and 1 mM PMSF were added to the cell suspension, and the mixture was vortexed for 2 min and then cooled on ice for 2 min (Choi et al, 2003).…”