Today, reactive oxygen molecules are no longer regarded by and large as culprits, since it has turned out that they are also involved in regulating major life processes such as growth and cell death. The right balance between oxidation and the reverse reaction, reduction, makes the difference between health and disease. "Oxidative stress" arises when this balance shifts towards oxidation-promoting processes.
So far, it has hardly been possible for scientists to measure the level of oxidation and, thus, the stress status of living cells. This will now be feasible thanks to a highly sensitive biomarker presented in the journal Nature Methods by Dr. Tobias Dick and co-workers of the German Cancer Research Center, jointly with colleagues from the University of Heidelberg.
The biosensor specifically measures the oxidation state of glutathione. This is an important protection molecule that captures a large portion of reactive oxygen molecules within a cell by oxidation. If much of a cell's glutathione is present in an oxidized state, this is an important indicator of the cell's overall oxidation level. The investigators equipped test cells with a fluorescent protein that reacts to changes in oxidation level by releasing light signals. Since the fluorescent protein on its own is not sensitive enough, it was coupled with an enzyme called glutaredoxin. This enzyme "measures" the oxidation state of glutathione and transmits the value to the fluorescent protein.
The stress biosensor developed by Dr. Dick and colleagues measures the slightest changes in the oxidation state of glutathione without destroying the cell. Even more relevant, however, is its precise time resolution, as Tobias Dick explained: "In order to measure short-term variations of oxidation state, the system needs to react instantly and dynamically. This is guaranteed with our biosensor, which works down to the scale of seconds." The measuring system allows researchers to determine those short-term variations that occur when reactive oxygen compounds are released as signaling molecules. However, the biosensor is equally suitable for use in pharmaceutical research, for example, to determine the effect of new substances or plant food constituents on oxidative processes and, thus, on the stress status of cells.
The task of the Deutsches Krebsforschungszentrum in Heidelberg - German Cancer Research Center, DKFZ - is to systematically investigate the mechanisms of cancer development and to identify cancer risk factors. The results of this basic research are expected to lead to new approaches in the prevention, diagnosis and treatment of cancer. The Center is financed to 90 percent by the Federal Ministry of Education and Research and to 10 percent by the State of Baden-Wuerttemberg. It is a member of the Helmholtz Association of National Research Centers - Helmholtz-Gemeinschaft Deutscher Forschungszentren e.V.