Transient light-induced intracellular oxidation revealed by redox biosensor.

Autor: Kolossov VL; Institute for Genomic Biology, University of Illinois at Urbana-Champaign, 1206 W. Gregory Drive, Urbana, IL 61801, USA. Electronic address: viadimer@illinois.edu., Beaudoin JN, Hanafin WP, DiLiberto SJ, Kenis PJ, Gaskins HR
Jazyk: angličtina
Zdroj: Biochemical and biophysical research communications [Biochem Biophys Res Commun] 2013 Oct 04; Vol. 439 (4), pp. 517-21. Date of Electronic Publication: 2013 Sep 08.
DOI: 10.1016/j.bbrc.2013.09.011
Abstrakt: We have implemented a ratiometric, genetically encoded redox-sensitive green fluorescent protein fused to human glutaredoxin (Grx1-roGFP2) to monitor real time intracellular glutathione redox potentials of mammalian cells. This probe enabled detection of media-dependent oxidation of the cytosol triggered by short wavelength excitation. The transient nature of light-induced oxidation was revealed by time-lapse live cell imaging when time intervals of less than 30s were implemented. In contrast, transient ROS generation was not observed with the parental roGFP2 probe without Grx1, which exhibits slower thiol-disulfide exchange. These data demonstrate that the enhanced sensitivity of the Grx1-roGFP2 fusion protein enables the detection of short-lived ROS in living cells. The superior sensitivity of Grx1-roGFP2, however, also enhances responsiveness to environmental cues introducing a greater likelihood of false positive results during image acquisition.
(Copyright © 2013 Elsevier Inc. All rights reserved.)
Databáze: MEDLINE
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