engleski

COMPARATIVE PROTEOME ANALYSIS OF YEAST'S ORGANELLES TREATED WITH LEAD OR IMIDACLOPRID

The yeast Saccharomyces cerevisiae is a useful model for studying the influence of different stress factors to the cells. It is known that metalinduced stress is mostly oxidative stress. The mechanism by which yeast cells combat the toxicity of pesticides is less known. The aim of this work was to analyse and compare the influence of pesticide imidacloprid and heavy metal lead to the proteome of Saccharomyces cerevisiae W303 yeast. During cultivation of the yeast biomass, defined amount of imidacloprid (0.2 mmol/L) or lead (10 ppm) was added to the growth media and their influence to the plasma membrane, mitochondrion, cytosol with microsomes and on the whole yeast cells has been monitored. Crude organelles were isolated after enzymatic disruption of the yeast’s cell wall. Proteins from the whole cells and isolated organelles were separated using 2D electrophoresis, stained using Mortz's silver staining method and processed using UVISpot program. Change in proteins expression was monitored through the change in their volume. Defined number of proteins was analysed using MS MALDITOF/TOF technique and identified using nrNCBI database. In the samples of cytosol with microsomes, plasma membrane and mitochondrion treated either with lead or imidacloprid, an increase in a total number of detected proteins, comparing nontreated samples, has occurred. No significant change in expression of proteins was observed in treated samples of plasma membrane while in treated samples of mitochondrion, enzyme succinate dehidrogenase shown significant increase in expression. In the samples of cytosol with microsomes, some proteins, like those involved in processes of carbohydrate metabolic pathways (particularly GAPDH) and enzyme Cu, Zn superoxide dismutase responsible for protection of yeast from oxidative stress, have increased their activity. The highest, significant increment in the expression in both treated samples of cytosol with microsomes, disulphide isomerase, the enzyme that catalyzes the formation and breakage of disulfide bonds between cysteine residues within proteins as they fold, was noticed. The involvement of Cu, Zn superoxide dismutase in yeast cell's response treated with imidacloprid imply the possibility that imidacloprid provoke oxidative stress in the yeast cells as the lead do.

proteome; Saccharomyces cerevisiae; imidacloprid; lead

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