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Aspectos estructurales para el reconocimiento de ATP por los receptores de ribonucleopéptido

Shun Nakano¹, Tsukasa Mashima, Akimasa Matsugami

¹Institute of Advanced Energy, Kyoto University, Japan.

Journal of the American Chemical Society

|March 5, 2011

Resumen

Los investigadores descubrieron una nueva estructura de ribonucleopéptido (RNP) para la creación de sensores moleculares. Este RNP se une al trifosfato de adenosina (ATP) mediante una única interacción triple U:A:U, lo que permite el reconocimiento molecular específico.

Área de la Ciencia:

Biología Molecular Biología Molecular
La bioquímica es la bioquímica.
Biología Estructural Biología estructural.

Sus antecedentes:

Los ribonucleopéptidos (RNP) ofrecen un marco modular para la construcción de receptores macromoleculares y sensores fluorescentes.
Los aptamers de ARN existentes para análogos de ATP exhiben diferentes secuencias de consenso que las identificadas RNP.

Objetivo del estudio:

Para dilucidar el mecanismo de unión al ATP de un nuevo RNP.
Para caracterizar la estructura tridimensional del complejo de unión RNP-sustrato.
Explorar el potencial de la ingeniería de la especificidad vinculante del RNP.

Principales métodos:

Espectroscopía de Resonancia Magnética Nuclear (RMN) Espectroscopía de Resonancia Magnética Nuclear (RMN) Espectroscopía de Resonancia Magnética Nuclear (RMN) Espectroscopía de Resonancia Magnética Nuclear (RMN) Espectroscopía de Resonancia Magnética Nuclear (RMN) Espectroscopía de Resonancia Magnética Nuclear (RMN) Espectroscopía de Resonancia Magnética Nuclear (RMN) Espectroscopía de Resonancia Magnética Nuclear (RMN).
Técnicas de mapeo enzimático y químico.
Estudios de mutación de nucleótidos del complejo RNP-adenosina.

Principales resultados:

Se identificó un RNP de unión al ATP con una secuencia de nucleótidos mínima para la unión al ATP.
Se determinó que el RNP interactúa con el anillo de adenina a través de una triple hélice U:A:U que involucra dos nucleótidos de uracilo invariantes.
Se demostró que este modo de reconocimiento difiere de los aptameros de ARN informados previamente para los derivados de ATP.
Demostró la conversión exitosa de la especificidad RNP de la unión de ATP a GTP utilizando un modo de reconocimiento triple C(+):G:C.

Conclusiones:

El complejo RNP-adenosina adopta un pliegue específico impulsado por triples U:A:U y emparejamiento de bases Hoogsteen A:U.
Este nuevo mecanismo de reconocimiento proporciona una base para la ingeniería de sensores moleculares basados en RNP con especificidades personalizadas.
La capacidad de cambiar la especificidad de unión de ATP a GTP destaca la versatilidad de este sistema RNP.

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