title: Mitochondria-Directed Fluorescent Probe for the Detection of Hydrogen Peroxide near Mitochondrial DNA.
abstract: It is important to detect hydrogen peroxide (H2O2) near mitochondrial DNA (mtDNA) because mtDNA is more prone to oxidative attack than nuclear DNA (nDNA). In this study, a mitochondria-targeted fluorescence probe, pep3-NP1, has been designed and synthesized. The probe contains a DNA-binding peptide, a H2O2 fluorescence reporter, and a positively charged red emissive styryl dye to facilitate accumulation in mitochondria. Due to groove binding of the peptide with DNA, the styryl dye of pep3-NP1 intercalated into the bases of DNA, leading to an increase in red fluorescence intensity (centered at 646 nm) and quantum yield. In this case, pep3-NP1 was a turn-on probe for labeling DNA. Subcellular locations of pep3-NP1 and MitoTracker suggested that pep3-NP1 mostly accumulated in the mitochondria of live cells. Namely, as an intracellular DNA marker, pep3-NP1 bound to mtDNA. In the presence of H2O2, pep3-NP1 emitted green fluorescence (centered at 555 nm). Thus, the ratio of green with red fluorescence of pep3-NP1 was suitable to reflect the change of the H2O2 level near mtDNA in living cells. The detecting limit for H2O2 was estimated at 2.9 and 5.0 μM in vitro and in cultured cells, respectively. The development of pep3-NP1 could help in studies to protect mtDNA from oxidative stress.
authors.affiliation: Department of Chemistry and Collaborative Innovation Center of Chemistry for Energy Materials, Fudan University , Shanghai 200433, China.
authors.forename: Ying
authors.initials: Y
authors.lastname: Wen
authors.affiliation: Department of Chemistry and Collaborative Innovation Center of Chemistry for Energy Materials, Fudan University , Shanghai 200433, China.
authors.forename: Keyin
authors.initials: K
authors.lastname: Liu
authors.affiliation: Department of Chemistry and Collaborative Innovation Center of Chemistry for Energy Materials, Fudan University , Shanghai 200433, China.
authors.forename: Huiran
authors.initials: H
authors.lastname: Yang
authors.affiliation: Department of Chemistry and Collaborative Innovation Center of Chemistry for Energy Materials, Fudan University , Shanghai 200433, China.
authors.forename: Yi
authors.initials: Y
authors.lastname: Liu
authors.affiliation: Department of Chemistry and Collaborative Innovation Center of Chemistry for Energy Materials, Fudan University , Shanghai 200433, China.
authors.forename: Liming
authors.initials: L
authors.lastname: Chen
authors.affiliation: Department of Chemistry and Collaborative Innovation Center of Chemistry for Energy Materials, Fudan University , Shanghai 200433, China.
authors.forename: Zhongkuan
authors.initials: Z
authors.lastname: Liu
authors.affiliation: Department of Chemistry and Collaborative Innovation Center of Chemistry for Energy Materials, Fudan University , Shanghai 200433, China.
authors.forename: Chunhui
authors.initials: C
authors.lastname: Huang
authors.affiliation: Department of Chemistry and Collaborative Innovation Center of Chemistry for Energy Materials, Fudan University , Shanghai 200433, China.
authors.forename: Tao
authors.initials: T
authors.lastname: Yi
mesh_terms: D002470:Cell Survival; D004272:DNA, Mitochondrial / Q000737:chemistry*; D005456:Fluorescent Dyes / Q000138:chemical synthesis / Q000737:chemistry*; D006367:HeLa Cells; D056945:Hep G2 Cells; D006801:Humans; D006861:Hydrogen Peroxide / Q000032:analysis*; D061986:MCF-7 Cells; D014407:Tumor Cells, Cultured
chemical_list: D004272:DNA, Mitochondrial; D005456:Fluorescent Dyes; D006861:Hydrogen Peroxide