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Resolución de la heterogeneidad conformacional específica del sitio en el reconocimiento molecular rico en prolina

Rachel E Horness¹, Edward J Basom¹, John P Mayer¹

¹Department of Chemistry, Indiana University , 800 East Kirkwood, Bloomington, Indiana 47405, United States.

Journal of the American Chemical Society

|January 20, 2016

Resumen

Este estudio revela cómo el dominio Sho1 SH3 reconoce a su compañero péptido Pbs2. Utilizando espectroscopia IR avanzada, los investigadores encontraron que el péptido adopta múltiples conformaciones al unirse, un proceso dinámico que se pierde en los métodos de RMN tradicionales.

Área de la Ciencia:

La bioquímica
Biología estructural
Espectroscopia

Sus antecedentes:

Los modelos de reconocimiento molecular de proteínas involucran cada vez más heterogeneidad y dinámica conformacional.
La caracterización experimental de estos procesos dinámicos sigue siendo un desafío importante.
El dominio de homología Src 3 (SH3) y sus ligandos peptídicos ricos en prolina (PR) son cruciales en la señalización celular.

Objetivo del estudio:

Caracterizar el reconocimiento molecular del dominio Sho1 SH3 con su secuencia Pbs2 PR.
Investigar los estados conformacionales adoptados por el péptido Pbs2 al unirse al dominio SH3.
Explorar la dinámica del reconocimiento molecular utilizando una combinación de espectroscopia IR y etiquetado isotópico.

Principales métodos:

Incorporación selectiva de enlaces carbono-deuterio (C-D) en el péptido Pbs2.
Espectroscopia infrarroja (IR) para el seguimiento de los cambios ambientales locales de los enlaces C-D.
Espectroscopia de resonancia magnética nuclear (RMN) para la comparación de la dinámica conformacional.
Caracterización de la formación del complejo peptídico SH3.

Principales resultados:

Los espectros IR de los péptidos Pbs2 etiquetados con C-D mostraron más absorciones de las esperadas tras la unión al dominio SH3, lo que indica estados de población múltiple.
Los espectros de RMN de péptidos etiquetados con (13) C en los mismos sitios exhibieron resonancias únicas, lo que sugiere una rápida interconversión en la escala de tiempo RMN.
Los datos proporcionan evidencia para el reconocimiento molecular de ajuste inducido que involucra estados conformacionales poblados transitoriamente.

Conclusiones:

El dominio Sho1 SH3 reconoce su péptido afín Pbs2 a través de un mecanismo de ajuste inducido.
Se poblan múltiples estados conformacionales durante el reconocimiento, que son demasiado transitorios para ser detectados por RMN convencional.
El enfoque combinado de espectroscopia IR y etiquetado CD ofrece un método poderoso para estudiar la dinámica rápida en el reconocimiento molecular.

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