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Aumento de la desnaturalización en frío de las proteínas en la interfaz hielo-agua

Andrea Arsiccio¹, James McCarty², Roberto Pisano¹

¹Department of Applied Science and Technology, Politecnico di Torino, 24 corso Duca degli Abruzzi, Torino 10129, Italy.

Journal of the American Chemical Society

|March 4, 2020

Resumen

La congelación de proteínas cerca de las interfaces hielo-agua provoca un rápido despliegue al bajar la barrera de energía. La adición de glucosa mitiga esta desnaturalización en frío actuando como un crioprotector.

Área de la Ciencia:

La biofísica
La bioquímica
Ciencias de los materiales

Sus antecedentes:

La estabilidad de las proteínas es crucial para el procesamiento biofarmacéutico, el almacenamiento y la criomicroscopia.
La interfaz hielo-agua durante la congelación es un factor desestabilizador conocido para las proteínas.
Los crioprotectores se usan a menudo para mitigar el daño de las proteínas inducido por la congelación.

Objetivo del estudio:

Investigar los mecanismos moleculares mediante los cuales la interfaz hielo-agua desestabiliza las proteínas durante la congelación.
Para entender el papel de los crioprotectores como la glucosa en la prevención del despliegue de proteínas a bajas temperaturas.

Principales métodos:

Utilizó simulaciones de dinámica molecular (MD) para modelar el comportamiento de las proteínas en la interfaz hielo-agua.
Analizó el paisaje de energía libre del despliegue de proteínas en presencia y ausencia de la interfaz.
Investigó el efecto de la glucosa en la hidratación y la estabilidad de las proteínas.

Principales resultados:

La interfaz hielo-agua reduce significativamente la barrera de energía libre para el despliegue de proteínas, lo que lleva a una rápida desnaturalización.
El despliegue de proteínas está mediado por moléculas de agua cercanas, no necesariamente por adsorción directa a la superficie del hielo.
Se demostró que la adición de glucosa mitiga la desnaturalización en frío a través de la exclusión preferente de las cadenas laterales de proteínas.

Conclusiones:

La interfaz hielo-agua es un potente impulsor del despliegue de proteínas, distinto de la simple desnaturalización en frío.
Comprender estos efectos interfaciales es crítico para optimizar las técnicas de criopreservación.
La glucosa protege eficazmente las proteínas alterando su caparazón de hidratación y reduciendo las interacciones interfaciales.

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