Open AccessBenzocoumarin Hydrazine: A Large Stokes Shift Fluorogenic Sensor for Detecting Carbonyls in Isolated Biomolecules and in Live Cells
Author(s) -
Kamalika Mukherjee,
Tak Ian Chio,
Han Gu,
Abhijit Banerjee,
Anthony M. Sorrentino,
Dan L. Sackett,
Susan Bane
Publication year - 2017
Publication title -
acs sensors
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 2.055
H-Index - 57
ISSN - 2379-3694
DOI - 10.1021/acssensors.6b00616
Subject(s) - biomolecule , fluorophore , stokes shift , hydrazone , fluorescence , chemistry , carbonylation , quenching (fluorescence) , photochemistry , nanotechnology , combinatorial chemistry , materials science , organic chemistry , biochemistry , catalysis , optics , physics , carbon monoxide
Detection and quantification of biomolecule carbonylation, a critical manifestation of oxidative stress, allows better understanding of associated disease states. Existing approaches for such analyses require further processing of cells and tissues, which leads to loss of both spatial and temporal information about carbonylated biomolecules in cells. Live cell detection of these species requires sensors that are nontoxic, sufficiently reactive with the biocarbonyl in the intracellular milieu, and detectable with commonly available instrumentation. Presented here is a new fluorescent sensor for biomolecule carbonyl detection: a hydrazine derivative of a benzocoumarin, 7-hydrazinyl-4-methyl-2H-benzo[h]chromen-2-one (BzCH), which meets these requirements. This probe is especially well suited for live cell studies. It can be excited by a laser line common to many fluorescence microscopes. The emission maximum of BzCH undergoes a substantial red shift upon hydrazone formation (from ∼430 to ∼550 nm), which is the result of fluorophore disaggregation. Additionally, the hydrazone exhibits an exceptionally large Stokes shift (∼195 nm). The latter properties eliminate self-quenching of the probe and the need to remove unreacted fluorophore for reliable carbonyl detection. Thus, biomolecule carbonylation can be detected and quantified in cells and in cell extracts in a one-step procedure using this probe.