title: A highly sensitive ratiometric fluorescent probe for the detection of cytoplasmic and nuclear hydrogen peroxide.
abstract: As a marker for oxidative stress and a second messenger in signal transduction, hydrogen peroxide (H2O2) plays an important role in living systems. It is thus critical to monitor the changes in H2O2 in cells and tissues. Here, we developed a highly sensitive and versatile ratiometric H2O2 fluorescent probe (NP1) based on 1,8-naphthalimide and boric acid ester. In response to H2O2, the ratio of its fluorescent intensities at 555 and 403 nm changed 1020-fold within 200 min. The detecting limit of NP1 toward H2O2 is estimated as 0.17 μM. It was capable of imaging endogenous H2O2 generated in live RAW 264.7 macrophages as a cellular inflammation response, and especially, it was able to detect H2O2 produced as a signaling molecule in A431 human epidermoid carcinoma cells through stimulation by epidermal growth factor. This probe contains an azide group and thus has the potential to be linked to various molecules via the click reaction. After binding to a Nuclear Localization Signal peptide, the peptide-based combination probe (pep-NP1) was successfully targeted to nuclei and was capable of ratiometrically detecting nuclear H2O2 in living cells. These results indicated that NP1 was a highly sensitive ratiometric H2O2 dye with promising biological applications.
authors.affiliation: Department of Chemistry and Concerted Innovation Center of Chemistry for Energy Materials, Fudan University , Shanghai 200433, China.
authors.forename: Ying
authors.initials: Y
authors.lastname: Wen
authors.forename: Keyin
authors.initials: K
authors.lastname: Liu
authors.forename: Huiran
authors.initials: H
authors.lastname: Yang
authors.forename: Yi
authors.initials: Y
authors.lastname: Li
authors.forename: Haichuang
authors.initials: H
authors.lastname: Lan
authors.forename: Yi
authors.initials: Y
authors.lastname: Liu
authors.forename: Xinyu
authors.initials: X
authors.lastname: Zhang
authors.forename: Tao
authors.initials: T
authors.lastname: Yi
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D001888:Boric Acids / Q000737:chemistry*; D045744:Cell Line, Tumor; D002467:Cell Nucleus / Q000187:drug effects / Q000378:metabolism* / Q000648:ultrastructure; D003593:Cytoplasm / Q000187:drug effects / Q000378:metabolism* / Q000648:ultrastructure; D004815:Epidermal Growth Factor / Q000494:pharmacology; D004847:Epithelial Cells / Q000187:drug effects / Q000378:metabolism / Q000648:ultrastructure; D004952:Esters; D005456:Fluorescent Dyes / Q000138:chemical synthesis / Q000737:chemistry*; D006801:Humans; D006861:Hydrogen Peroxide / Q000032:analysis*; D057230:Limit of Detection; D008264:Macrophages / Q000187:drug effects / Q000378:metabolism / Q000648:ultrastructure; D051379:Mice; D008856:Microscopy, Fluorescence; D008969:Molecular Sequence Data; D053644:Naphthalimides / Q000737:chemistry*; D019913:Nuclear Localization Signals / Q000737:chemistry; D018384:Oxidative Stress
chemical_list: D001888:Boric Acids; D004952:Esters; D005456:Fluorescent Dyes; D053644:Naphthalimides; D019913:Nuclear Localization Signals; D004815:Epidermal Growth Factor; D006861:Hydrogen Peroxide