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Escisión de enlaces peptídicos regulados por conformación en la bradicinina

Daniel R Fuller¹, Christopher R Conant¹, Tarick J El-Baba¹

¹Department of Chemistry , Indiana University , Bloomington , Indiana 47405 , United States.

Journal of the American Chemical Society

|July 21, 2018

Resumen

La bradicinina (BK) se somete a una escisión no enzimática única en el enlace Pro2-Pro3, provocada por la protonización e isomerización inducidas por la temperatura. Esta vía intrínseca, resistente a las enzimas humanas, ofrece información sobre el procesamiento de péptidos y la antigenización.

Área de la Ciencia:

La bioquímica
Física Química
Química analítica

Sus antecedentes:

La bradicinina (BK) es un péptido con funciones biológicas conocidas.
Comprender la estabilidad y el procesamiento de los péptidos es crucial para el desarrollo de fármacos y los estudios biológicos.
Las vías de escisión de péptidos intrínsecos pueden ofrecer información sobre los mecanismos de resistencia biológica.

Objetivo del estudio:

Investigar las estabilidades conformacionales de diferentes conformaciones de bradicininas (BK) utilizando movilidad iónica y espectrometría de masas.
Para aclarar el mecanismo de una nueva división de enlace Pro2-Pro3 no enzimática en BK.
Determinar los parámetros termodinámicos de los estados de transición involucrados en el procesamiento de BK.

Principales métodos:

Espectrometría de movilidad iónica junto con espectrometría de masas (IM-MS).
Estudios cinéticos dependientes de la temperatura.
Análisis de las reacciones de protonación y cambios conformacionales.

Principales resultados:

Las temperaturas elevadas inducen una reacción de protonación lenta ([BK+2H]2+ → [BK+3H]3+) regulada por la isomerización trans → cis de Arg1-Pro2.
La conformación all-cis [BK+3H]3+ rompe espontáneamente el enlace Pro2-Pro3 con alta especificidad.
Esta escisión se produce a través de múltiples intermedios, regulados por la isomerización Arg1-Pro2, y es biológicamente resistente.
Se determinó la termoquímica del estado de transición para la protonación y la escisión de enlaces.

Conclusiones:

Se identifica una nueva vía de escisión no enzimática para la bradicinina en el enlace Pro2-Pro3.
Esta vía está regulada por la isomerización Arg1-Pro2 trans → cis y la protonación.
La resistencia biológica a esta vía de procesamiento intrínseco tiene implicaciones para la antigenicidad del péptido.

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