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La unión no específica del tripolifosfato de adenosina y el tripolifosfato modula el comportamiento de fase de la
Matja Zalar1, Jordan Bye1, Robin Curtis1
1Manchester Institute of Biotechnology, Department of Chemical Engineering, Faculty of Science and Engineering, The University of Manchester, 131 Princess Street, ManchesterM1 7DN, U.K.
Journal of the American Chemical Society
|January 6, 2023
Resumen
El tripolifosfato de adenosina (ATP) se une de manera no específica a las proteínas como la lisozima, influyendo en la solubilidad y agregación de las proteínas. Esta interacción, principalmente a través del grupo polifosfato, tiene implicaciones para la cristalización de proteínas y los procesos celulares.
Área de la Ciencia:
- La bioquímica
- Biología estructural
- La biofísica
Sus antecedentes:
- Se sabe que el tripolifosfato de adenosina (ATP) interactúa con las proteínas, afectando la formación de condensado biomolecular y la agregación de proteínas.
- La naturaleza precisa de las interacciones proteína-ATP no específicas y su impacto en la solubilidad de las proteínas siguen sin caracterizarse en gran medida.
Objetivo del estudio:
- Para aclarar las características detalladas de unión de ATP a una proteína modelo, la lisozima.
- Investigar los efectos de la unión de ATP en la solubilidad, agregación y cristalización de las proteínas.
Principales métodos:
- Se empleó la cristalografía de rayos X para determinar la base estructural de la unión de ATP.
- Se utilizó la espectroscopia de Resonancia Magnética Nuclear (RMN) para identificar los sitios de unión al ATP y caracterizar las afinidades de unión.
- Se realizaron estudios de interacción proteína-proteína en la fase de solución con diferentes concentraciones de sal y en presencia de magnesio.
Principales resultados:
- Se identificaron seis sitios de unión de ATP en la lisozima: un sitio conocido de alta afinidad y cinco nuevos sitios no específicos con afinidades millimolares que también se unen al tripolifosfato (TPP).
- La unión de ATP, principalmente a través de su fracción de polifosfato, se produce preferentemente en los residuos de arginina sobre la lisina en sitios no específicos.
- El ATP y el TPP mostraron efectos similares en las interacciones proteína-proteína, induciendo la precipitación a bajas concentraciones de sal y la redisolución a concentraciones más altas.
- La adición de magnesio al ATP no afectó la afinidad de unión, pero evitó la precipitación de lisozima.
Conclusiones:
- El ATP se une no específicamente a las proteínas plegadas bajo condiciones fisiológicas, principalmente a través de su grupo polifosfato.
- La unión de ATP influye en la solubilidad de las proteínas y puede prevenir la agregación de proteínas, con mecanismos potenciales identificados.
- Los hallazgos ofrecen información sobre las estrategias de cristalización de proteínas y el papel del ATP en la homeostasis de proteínas celulares.
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