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Zawartość zarchiwizowana w dniu 2024-06-10

Regulation of mammalian iron metabolism by oxidative stress

Cel



In the proposed research plan, we intend to study physiological and mechanistic aspects of the response of IRP-1 to oxidative stress. By employing different agents which cause oxidative stress to cells, we will compare the effects of extracellular and intracellular oxidative stress on IRP-1. For this, we need to measure extra- and intracellular H2o2 levels. A highly sensitive chemiluminescent assay will be applied for the determination of H2o2 in the tissue culture supernatant (Mueller and Arnhold, 1995), while intracellular levels of H2o2 will be assessed by FACS analysis, using a fluorescent redox- sensitive probe. These tools will enable a detailed characterisation of the effects of H2r2 on IRP-1. In our previous work, we have provided evidence that extracellular H2o2 causes an accelerated turnover of the 4Fe-4S cluster, possibly via a signaling pathway. We now propose to further address this hypothesis, aiming to elucidate the underlying mechanism. This would require to establish an in vitro system to recapitulate the effects observed in intact cells. Such a system would enable us to dissect the early steps of the H202-response from the actual removal of the 4Fe-4S cluster. Furthermore it would provide a framework to isolate and biochemically characterize possible cluster- destabilizing activities. Training, content (objective, benefit and expected impact) The goals of this project lie within the interests of the host lab. The choise of this host lab is appropriate, as my previous training at EMBL on modern techniques of molecular biology will be meaningfully complemented by the expertise and know-how of Dr. Papavassiliou's lab. In addition, I believe that I could contribute with my knowledge in the undergraduate as well as graduate programmes of the Medical School of Patras.

Dziedzina nauki (EuroSciVoc)

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Koordynator

Aristotle University of Thessaloniki
Wkład UE
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Adres
Analytical Chemistry Lab
54006 Thessaloniki
Grecja
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