A tautomeric zinc sensor for ratiometric fluorescence imaging: Application to nitric oxide-induced release of intracellular zinc
AUTOR(ES)
Chang, Christopher J.
FONTE
National Academy of Sciences
RESUMO
Zinc is an essential metal ion for human growth and development, the disruption of cellular Zn2+ homeostasis being implicated in several major disorders including Alzheimer's disease, diabetes, and cancer. The molecular mechanisms of Zn2+ physiology and pathology are insufficiently understood, however, owing in part to the lack of tools for measuring changes in intracellular Zn2+ concentrations with high spatial and temporal fidelity. To address this critical need, we have synthesized, characterized, and applied an intracellular fluorescent probe for the ratiometric imaging of Zn2+ based on a tautomeric seminaphthofluorescein platform. Zin-naphthopyr 1 (ZNP1) affords single-excitation, dual-emission ratiometric detection of intracellular Zn2+ through Zn2+-controlled switching between fluorescein and naphthofluorescein tautomeric forms. The probe features visible excitation and emission profiles, excellent selectivity responses for Zn2+ over competing Ca2+ and Mg2+ ions at intracellular concentrations, a dissociation constant (Kd) for Zn2+ of <1 nM, and an 18-fold increase in fluorescence emission intensity ratio (λ624/λ528) upon zinc binding. We demonstrate the value of the ZNP1 platform for biological applications by imaging changes in intracellular [Zn2+] in living mammalian cells. Included is the ratiometric detection of endogenous pools of intracellular Zn2+ after NO-induced release of Zn2+ from cellular metalloproteins. We anticipate that ZNP1 and related probes should find utility for interrogating the biology of Zn2+.
ACESSO AO ARTIGO
http://www.pubmedcentral.nih.gov/articlerender.fcgi?artid=337018Documentos Relacionados
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