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Photo-CIDNP revela diferencias en la compactación de los estados no nativos de la lisozima

Christian Schlörb¹, Sarah Mensch, Christian Richter

¹Institute for Organic Chemistry and Chemical Biology, Center for Biomolecular Magnetic Resonance, Johann Wolfgang Goethe-University, Marie-Curie-Strasse 11, D-60439 Frankfurt/M, Germany.

Journal of the American Chemical Society

|February 9, 2006

Resumen

La polarización nuclear dinámica inducida fotoquímicamente (foto-CIDNP) revela cambios estructurales en las proteínas. Este método mapea la exposición al disolvente y la dinámica de los residuos de triptófano en la lisozima de clara de huevo de gallina.

Área de la Ciencia:

Química biofísica y bioquímica.
Espectroscopia de RMN de proteínas Espectroscopia de RMN de proteínas
Biología Estructural Biología estructural.

Sus antecedentes:

Los estados proteicos no nativos presentan conjuntos estructurales complejos.
Comprender la dinámica de las proteínas y la accesibilidad de los disolventes es crucial para caracterizar estos estados.
Los residuos de triptófano son valiosas sondas debido a sus propiedades espectroscópicas únicas.

Objetivo del estudio:

Para caracterizar los conjuntos estructurales de la lisozima de la clara de huevo de gallina no nativa.
Para investigar la exposición al disolvente y la dinámica de los residuos individuales de triptófano.
Para correlacionar los conocimientos estructurales con el agrupamiento hidrofóbico utilizando foto-CIDNP.

Principales métodos:

Se emplearon experimentos bidimensionales de foto-CIDNP.
Se utilizaron espectros HSQC convencionales para la comparación.
Las mediciones de relajación transversal analizaron la dinámica de NH de la cadena vertebral y lateral.

Principales resultados:

Se determinó la accesibilidad diferencial al disolvente de seis residuos de triptófano en la lisozima no nativa.
La accesibilidad del disolvente correlacionada con la dinámica de los residuos.
El agrupamiento hidrofóbico alrededor de los residuos de triptófano fue respaldado por datos, incluido un mutante W62G.

Conclusiones:

Photo-CIDNP es eficaz para la caracterización estructural de conjuntos de proteínas no nativas.
El estudio proporciona información sobre las propiedades dinámicas y estructurales de la lisozima.
El agrupamiento hidrofóbico juega un papel importante en el comportamiento observado de los residuos de triptófano.

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