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¿Cómo desestabilizan los detergentes no iónicos de cadena corta los receptores acoplados a la proteína G?

Sangbae Lee¹, Allen Mao¹, Supriyo Bhattacharya¹

¹Department of Molecular Immunology, Beckman Research Institute of the City of Hope , 1500 E. Duarte Road, Duarte, California 91010, United States.

Journal of the American Chemical Society

|October 30, 2016

Resumen

Los detergentes de cadena corta como el octilglucósido desestabilizan los receptores acoplados a la proteína G (GPCR) alterando su estructura. Los detergentes más suaves podrían mejorar la purificación y cristalización de GPCR.

Área de la Ciencia:

La bioquímica
Biología molecular
Biofísica computacional

Sus antecedentes:

Los receptores acoplados a proteínas G (GPCR) requieren solubilización con detergente para su purificación.
Se sabe que los detergentes de cadena corta son más desnaturalizantes que los detergentes de cadena larga para los GPCR.
Los mecanismos moleculares subyacentes a la desestabilización GPCR inducida por detergente no se comprenden completamente.

Objetivo del estudio:

Investigar las bases moleculares de la desestabilización del detergente en los GPCR.
Comprender cómo los diferentes detergentes afectan la integridad estructural de los GPCR.
Identificar estrategias para el desarrollo de detergentes más suaves para la investigación GPCR.

Principales métodos:

Se emplearon simulaciones de dinámica molecular atómica.
Las simulaciones se centraron en los mutantes termostabilizados del receptor de adenosina (A2A R).
Los receptores fueron modelados dentro de las bicapas lipídicas y las micelas de detergente (alquilmaltosidos y alquilglucósidos).

Principales resultados:

El octilglucósido y el nonilglucósido alteraron la estructura R de A2A, causando una disminución de la α-helicidad y el empaque interhelical.
Las moléculas de detergente penetraron en el núcleo transmembrana del receptor en detergentes duros.
El hemisuccinato de colesterol estabilizó el receptor en el dodecilmaltosido mejorando las propiedades de empaque y micela.
Se aclaró un proceso de desestabilización en tres etapas por octilglucósido.

Conclusiones:

La estructura del detergente, específicamente la longitud de la cadena de alquilo y la hidratación del grupo principal, influye en la estabilidad del GPCR.
Los detergentes más suaves, potencialmente basados en octilglucósido, podrían mejorar la purificación y cristalización de GPCR.
La comprensión de las interacciones detergente-micela-receptor es clave para preservar la estructura y la función de las GPCR.

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