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Oligopeptide Competition Assay for Phosphorylation Site Determination
Published on: May 18, 2017
Detección selectiva de los inhibidores alostéricos de la fosfatasa
Ralf Schneider1, Claudia Beumer, Jeffrey R Simard
1Chemical Genomics Centre der Max-Planck-Gesellschaft, Dortmund, Germany.
Journal of the American Chemical Society
|April 25, 2013
Resumen
El desarrollo de nuevos inhibidores de la fosfatasa es crucial para el tratamiento de enfermedades como el cáncer. Este estudio introduce un nuevo ensayo de cribado de alto rendimiento para descubrir inhibidores alostéricos de la fosfatasa, superando las limitaciones de los métodos anteriores.
Área de la Ciencia:
- La bioquímica es la bioquímica.
- Enzimología Enzimología.
- Descubrimiento de Drogas Descubrimiento de Drogas
Sus antecedentes:
- La fosforilación de proteínas por kinasas y fosfatasas regula las funciones celulares.
- La desregulación de la fosforilación contribuye a enfermedades como el cáncer y la inflamación.
- Si bien los inhibidores de la quinasa son exitosos, ningún inhibidor de la fosfatasa está clínicamente aprobado debido a problemas con la selectividad y la carga.
Objetivo del estudio:
- Desarrollar un nuevo ensayo para el descubrimiento de inhibidores alostéricos de la fosfatasa.
- Para hacer frente a la falta de sistemas de cribado adecuados para los moduladores alostéricos.
- Permitir la identificación de fármacos candidatos con propiedades farmacológicas mejoradas.
Principales métodos:
- Desarrollo de un nuevo ensayo vinculante.
- Prueba diseñada para detectar ligandos que se unen a un bolsillo alostérico.
- Prueba validada para el cribado de alto rendimiento (HTS).
Principales resultados:
- El ensayo detecta con éxito los ligandos que se unen a la bolsa alostérica de la proteína tirosina fosfatasa 1B (PTP1B).
- El ensayo demuestra una clara discriminación entre los ligandos alostéricos y substrato-competitivos.
- El ensayo es adecuado para aplicaciones de cribado de alto rendimiento.
Conclusiones:
- Se ha desarrollado un nuevo ensayo selectivo para el descubrimiento del inhibidor de la fosfatasa alostérica.
- Este ensayo facilita la identificación de nuevos moduladores de la fosfatasa.
- El ensayo supera las limitaciones de los métodos anteriores, allanando el camino para nuevas terapias.
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