Development of a General Aza-Cope Reaction Trigger Applied to Fluorescence Imaging of Formaldehyde in Living Cells

• Autores: Bruemmer, Kevin J.; Walvoord, Ryan R.; Brewer, Thomas F.; Burgos Barragan, Guillermo; Wit, Niek; Pontel, Lucas Blas; Patel, Ketan J.; Chang, Christopher J.
• Año de publicación: 2017
• Idioma: inglés
• Tipo de recurso: artículo
• Materia: FORMALDEHYDE; AZA-COPE REACTION; CELLS; ENDOGENOUS
• Institución: Consejo Nacional de Investigaciones Científicas y Técnicas

Formaldehyde (FA) is a reactive signaling molecule that is continuously produced through a number of central biological pathways spanning epigenetics to one-carbon metabolism. On the other hand, aberrant, elevated levels of FA are implicated in disease states ranging from asthma to neurodegenerative disorders. In this context, fluorescence-based probes for FA imaging are emerging as potentially powerful chemical tools to help disentangle the complexities of FA homeostasis and its physiological and pathological contributions. Currently available FA indicators require direct modification of the fluorophore backbone through complex synthetic considerations to enable FA detection, often limiting the generalization of designs to other fluorophore classes. To address this challenge, we now present the rational, iterative development of a general reaction-based trigger utilizing 2-aza-Cope reactivity for selective and sensitive detection of FA in living systems. Specifically, we developed a homoallylamine functionality that can undergo a subsequent self-immolative β-elimination, creating a FA-responsive trigger that is capable of masking a phenol on a fluorophore or any other potential chemical scaffold for related imaging and/or therapeutic applications. We demonstrate the utility of this trigger by creating a series of fluorescent probes for FA with excitation and emission wavelengths that span the UV to visible spectral regions through caging of a variety of dye units. In particular, Formaldehyde Probe 573 (FAP573), based on a resorufin scaffold, is the most red-shifted and FA sensitive in this series in terms of signal-to-noise responses and enables identification of alcohol dehydrogenase 5 (ADH5) as an enzyme that regulates FA metabolism in living cells. The results provide a starting point for the broader use of 2-aza-Cope reactivity for probing and manipulating FA biology.
Fil: Bruemmer, Kevin J.. University of California at Berkeley; Estados Unidos
Fil: Walvoord, Ryan R.. University of California at Berkeley; Estados Unidos
Fil: Brewer, Thomas F.. University of California at Berkeley; Estados Unidos
Fil: Burgos Barragan, Guillermo. Medical Research Council. Laboratory of Molecular Biology; Reino Unido
Fil: Wit, Niek. Medical Research Council. Laboratory of Molecular Biology; Reino Unido
Fil: Pontel, Lucas Blas. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Parque Centenario. Instituto de Investigación en Biomedicina de Buenos Aires - Instituto Partner de la Sociedad Max Planck; Argentina. Medical Research Council. Laboratory of Molecular Biology; Reino Unido
Fil: Patel, Ketan J.. University of Cambridge; Reino Unido. Medical Research Council. Laboratory of Molecular Biology; Reino Unido
Fil: Chang, Christopher J.. University of California at Berkeley; Estados Unidos