title: A specific fluorescence probe for hydrogen peroxide detection in peroxisomes.
abstract: Hydrogen peroxide is an important mediator in cell signalling and cell death. Apart from the mitochondrion the peroxisome is the most important cellular site for the generation and scavenging of hydrogen peroxide. Peroxisomes contain various oxidases, e.g. for the metabolism of long-chain fatty acids, polyamines, and for the oxidation of urate, which form hydrogen peroxide. Widely-used chemical probes for the detection of hydrogen peroxide like dichlorofluorescein diacetate (DCFDA) often lack in specificity and the possibility of compartment-specific measurement. To overcome these disadvantages, Belousov et al. developed the novel hydrogen peroxide sensitive fluorescent protein HyPer. In the present study the HyPer protein was fused with the PTS1 tag for a specific hydrogen peroxide detection in peroxisomes. The localization of the HyPer protein in the peroxisomes was confirmed by immunofluorescence and the functionality by fluorescence microscopy and flow cytometry analyses. The presented HyPer-Peroxi fluorescent protein is a valuable tool for studying hydrogen peroxide generation within the peroxisomes.
authors.affiliation: Institute of Clinical Biochemistry, Hannover Medical School, 30623 Hannover, Germany.
authors.forename: Wiebke
authors.initials: W
authors.lastname: Gehrmann
authors.forename: Matthias
authors.initials: M
authors.lastname: Elsner
mesh_terms: D000818:Animals; D002374:Catalase / Q000378:metabolism; D045744:Cell Line, Tumor; D005453:Fluorescence; D006861:Hydrogen Peroxide / Q000032:analysis* / Q000378:metabolism; D008928:Mitochondria / Q000378:metabolism; D015335:Molecular Probes / Q000032:analysis* / Q000378:metabolism; D010084:Oxidation-Reduction; D010088:Oxidoreductases / Q000378:metabolism; D020675:Peroxisomes / Q000737:chemistry* / Q000378:metabolism; D051381:Rats; D011994:Recombinant Proteins / Q000378:metabolism*; D013379:Substrate Specificity; D014527:Uric Acid / Q000378:metabolism
chemical_list: D015335:Molecular Probes; D011994:Recombinant Proteins; D014527:Uric Acid; D006861:Hydrogen Peroxide; D010088:Oxidoreductases; D002374:Catalase