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Intein Zymogens: ensamblaje condicional y escisión de inteínas divididas a través de la proteólisis dirigida

Josef A Gramespacher¹, Adam J Stevens¹, Duy P Nguyen²

¹Frick Laboratory, Department of Chemistry, Princeton University , Princeton, New Jersey 08544, United States.

Journal of the American Chemical Society

|June 1, 2017

Resumen

Los investigadores diseñaron inteínas divididas para el transesplicado de proteínas (PTS) que se activan en presencia de proteasa. Esta innovación permite nuevos sensores y actuadores de proteínas que responden a estímulos como patógenos y luz.

Área de la Ciencia:

Biología Química
Biología molecular
Biotecnología

Sus antecedentes:

Las inteínas divididas naturalmente facilitan el transesplicado de proteínas (PTS) para la ligadura polipeptídica.
Las aplicaciones actuales de los inteles divididos son limitadas en el control condicional.

Objetivo del estudio:

Para diseñar inteínas divididas condicionales activadas por proteasas definidas por el usuario.
Desarrollar nuevos sensores y actuadores de proteínas utilizando este sistema.

Principales métodos:

Los "zimógenos" de proteínas se diseñaron para asociarse solo en presencia de una proteasa específica.
Se comprobó la compatibilidad del sistema con diversos estímulos (patógenos, luz).
Para la multiplexación se utilizaron pares de inteín divididos ortogonales.

Principales resultados:

Se ha demostrado la transmisión condicional de proteínas por proteasa (PTS).
Se han creado con éxito sensores y actuadores de proteínas que responden a diversos estímulos.
Mostró el potencial de los sistemas de sensores multiplexados que utilizan inteños ortogonales.

Conclusiones:

Las células divididas condicionales ofrecen una plataforma versátil para nuevos biosensores y actuadores.
Este sistema activado por la proteasa mejora el control sobre la ligadura de proteínas.
La tecnología apoya el desarrollo de sistemas avanzados de detección biológica multiplexados.

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