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Las interacciones dinámicas de proteínas multicorpos sugieren vías versátiles para el tráfico de cobre
Aaron M Keller1, Jaime J Benítez, Derek Klarin
1Department of Chemistry and Chemical Biology, Cornell University, Ithaca, New York 14853, USA.
Journal of the American Chemical Society
|May 15, 2012
Resumen
El chaperón de cobre humano Hah1 le entrega el cobre a Wilson.
Área de la Ciencia:
- Bioquímica y Biología Molecular.
- Mecanismos de transporte de cobre intracelular Mecanismos de transporte de cobre intracelular Mecanismos de transporte de cobre intracelular Mecanismos de transporte de cobre intracelular
Sus antecedentes:
- El chaperón de cobre humano Hah1 facilita la entrega intracelular de cobre (Cu +) a la proteína de la enfermedad de Wilson (WDP).
- WDP posee seis dominios de unión de metales (MBD) que reciben Cu+ de Hah1, pero el papel de múltiples MBD en el tráfico de cobre sigue sin estar claro.
- Investigaciones anteriores exploraron las interacciones de Hah1 con MBDs WDP aislados.
Objetivo del estudio:
- Para investigar la dinámica de interacción entre Hah1 y una construcción WDP de doble dominio (MBD34).
- Para entender cómo Hah1 interactúa con múltiples MBDs dentro del sistema multidominio WDP.
- Para aclarar la importancia funcional de la multiplicidad de MBD en el tráfico de cobre.
Principales métodos:
- Empleado transferencia de energía de resonancia de fluorescencia de una sola molécula (smFRET) combinado con la captura de vesículas.
- Probe sistemáticamente las dinámicas de interacción de Hah1-MBD3, Hah1-MBD4 y MBD3-MBD4 intramoleculares.
- Utilizó posiciones alternas de donante y aceptador para analizar geometrías de interacción.
Principales resultados:
- Geometrías de interacción de interconversión conservadas observadas tanto en las interacciones intermoleculares Hah1-MBD como en las intramoleculares MBD-MBD.
- Se encontró que las interacciones Hah1-MBD dentro de la construcción MBD34 se estabilizan por un orden de magnitud en comparación con los MBD aislados.
- La evidencia termodinámica y cinética recopilada indica que Hah1 puede interactuar simultáneamente con MBD3 y MBD4.
Conclusiones:
- La mayor estabilidad de interacción de Hah1 con el sistema multi-MBD sugiere un mecanismo de entrega de cobre más robusto.
- Las interacciones dinámicas intramoleculares MBD-MBD contribuyen a la versatilidad del tráfico de cobre.
- La capacidad de Hah1 para interactuar simultáneamente con múltiples MBD de WDP apunta a una vía de transporte de cobre de Hah1 a WDP eficiente y adaptable.
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