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Estructura cristalina del triángulo de origami de proteínas en espiral diseñado de Novo

Tadej Satler^1,2, San Hadži^1,3, Roman Jerala^1,4

¹Department of Synthetic Biology and Immunology, National Institute of Chemistry, 1000 Ljubljana, Slovenia.

Journal of the American Chemical Society

|July 24, 2023

Resumen

El origami de proteínas en espiral (CCPO) permitió la creación de nuevas estructuras de proteínas triangulares. La estructura cristalina revela un nudo de proteína de tipo trébol, que valida los principios de diseño del CCPO.

Área de la Ciencia:

Biología estructural
Diseño de las proteínas
La bioquímica

Sus antecedentes:

El origami de proteínas en espiral (CCPO) es una estrategia para diseñar nuevas estructuras de proteínas utilizando bloques de construcción en espiral modulares.
Los diseños anteriores de CCPO carecían de datos estructurales de alta resolución, lo que limitaba la comprensión de su estabilidad y plegamiento.
Las proteínas naturales suelen adoptar pliegues globulares, mientras que CCPO apunta a arquitecturas poliédricas.

Objetivo del estudio:

Para determinar la estructura cristalina de alta resolución de un CCPO de una sola cadena diseñado en forma triangular.
Proporcionar conocimientos moleculares sobre los mecanismos de estabilización de las estructuras CCPO.
Validar la estrategia de diseño modular de proteínas en espiral.

Principales métodos:

Diseño de origami de proteínas en espiral (CCPO) de una proteína triangular.
Los ensayos de cristalización, incluida la ciclización y la adición de nanobody, finalmente tuvieron éxito con la inclusión de homodímero GCN2.
Cristalografía de rayos X para determinar la estructura tridimensional de la proteína.
Análisis de las interacciones entre los segmentos y el empaque de cristales.

Principales resultados:

La estructura cristalina de un triángulo CCPO de una sola cadena se determinó con alta resolución.
La estructura triangular está formada por dímeros enrollados paralelos ortogonales (P1: P2, P3: P4, GCN2) conectados por enlaces.
El pliegue de la proteína forma un nudo topológico de tipo trébol, que no fue predicho con precisión por AlphaFold2.
Las interacciones entre los segmentos en los vértices y el embalaje de cristal denso estabilizan la estructura.

Conclusiones:

La estructura cristalina determinada valida la estrategia de diseño del CCPO para crear nuevas topologías de proteínas.
Se obtuvieron conocimientos moleculares sobre la estabilización de las estructuras de CCPO, destacando el papel de las interacciones de vértices y el empaque de cristales.
Los hallazgos abren nuevas vías para diseñar arquitecturas complejas de proteínas y comprender las limitaciones del plegamiento de proteínas.

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