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El desbloqueo del dominio de transporte establece la tasa de absorción de un transportador de aspartato
Nurunisa Akyuz1, Elka R Georgieva2, Zhou Zhou1
1Department of Physiology and Biophysics, Weill Cornell Medical College, Cornell University, 1300 York Avenue, New York, New York 10065, USA.
Nature
|February 6, 2015
Resumen
Los transportadores de glutamato despejan los neurotransmisores. Este estudio visualiza sus resultados.
Área de la Ciencia:
- Biología Estructural Biología estructural.
- La bioquímica es la bioquímica.
- La neurociencia es la neurociencia.
Sus antecedentes:
- Los transportadores de glutamato son cruciales para la terminación de la neurotransmisión y la prevención de la excitotoxicidad.
- Estudios cristalográficos anteriores revelaron un andamio trímero y dominios de transporte móviles en GltPh.
- El mecanismo de la translocación del sustrato por estos transportadores no estaba claro.
Objetivo del estudio:
- Para observar directamente los movimientos dinámicos de los dominios del transportador de glutamato.
- Para establecer una relación entre la dinámica del dominio del transporte y la tasa de transporte del sustrato.
- Investigar el efecto de las mutaciones "humanizadoras" en la dinámica y el mecanismo del transportador.
Principales métodos:
- Imágenes de transferencia de energía por resonancia de fluorescencia de molécula única (smFRET).
- Reconstitución de proteoliposomas con GltPh purificado.
- Utilizando gradientes fisiológicos de iones.
- Análisis cristalográfico y computacional.
Principales resultados:
- Visualización directa de los movimientos del dominio de transporte "como en un ascensor" en GltPh.
- El GltPh mutado (con características humanas) mostró un aumento en la dinámica del dominio.
- El aumento de la dinámica se correlacionó directamente con una mayor velocidad de transporte del sustrato.
- Las mutaciones favorecían los estados intermedios desbloqueados con una mayor exposición al solvente.
Conclusiones:
- Las dinámicas del dominio de transporte están directamente acopladas a la translocación del sustrato en los transportadores de glutamato.
- Las mutaciones "humanizantes" mejoran la eficiencia del transportador al promover estados dinámicos y desbloqueados.
- Esto proporciona un vínculo mecánico entre la estructura, la dinámica y la función en el transporte de neurotransmisores.
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