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Una plataforma generalizable para interrogar las consecuencias específicas del objetivo y de la señal de las

Hong-Yu Lin¹, Joseph A Haegele¹, Michael T Disare¹

¹†Department of Chemistry and Chemical Biology, Cornell University, Ithaca, New York 14853, United States.

Journal of the American Chemical Society

|April 25, 2015

Resumen

Desarrollamos T-REX, una caja de herramientas dirigida a electrófilos reactivos y oxidantes, para modificar con precisión proteínas específicas. Este método permite cuantificar los cambios en la señalización celular de las modificaciones electrofílicas específicas, incluso cuando fallan los enfoques generales.

Área de la Ciencia:

Biología Química Biología química.
La señalización molecular.
La proteómica es la proteómica.

Sus antecedentes:

Los electrófilos de pequeñas moléculas interactúan con muchas proteínas activas de cisteína.
Los efectos biológicos son a menudo específicos del quimiotipo, lo que dificulta el estudio de objetivos individuales.

Objetivo del estudio:

Desarrollar un método para modificar con precisión objetivos intracelulares específicos con electrófilos.
Para cuantificar el impacto de estas modificaciones específicas en las vías de señalización aguas abajo.

Principales métodos:

Construyó una plataforma para entregar diversos electrofilos de señalización derivados de lípidos a proteínas específicas.
Investigada la eficiencia de la focalización, encontrar la tasa de alquilación es crucial.
Evaluación de las consecuencias de la señalización aguas abajo de las modificaciones específicas de la proteína objetivo y de la señal de las pequeñas moléculas.

Principales resultados:

T-REX permite el análisis cuantitativo de la activación de la señalización del elemento de respuesta antioxidante (ARE) dependiente del factor de transcripción Nrf2 a través de la modificación de Keap1.
Keap1 actúa como un sensor promiscuo, respondiendo de manera similar a diferentes señales electrofílicas para la inducción de Nrf2-ARE.
T-REX indujo con éxito la activación celular donde la amplia aplicación de electrófilos falló debido a la citotoxicidad.

Conclusiones:

T-REX proporciona una herramienta novedosa para elucidar las consecuencias funcionales de modificaciones electrofílicas específicas.
La plataforma permite estudiar eventos de señalización dirigidos en un contexto celular no perturbado.
Esta metodología abre nuevas vías para comprender las respuestas celulares complejas a las señales químicas.

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