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La estructura cristalina de rayos X de BorF, la subunidad de flavina reductasa de una triptófano halogenasa

Zheng Ma¹, Emily W Rady¹, Aravinda J de Silva¹

¹Department of Chemistry and Biochemistry, College of Natural Sciences and Mathematics, The University of Toledo, 2801 W. Bancroft St, Toledo, OH, 43606, U.S.A.

bioRxiv : the preprint server for biology

|August 20, 2025

Resumen

La flavinreductasa BorF utiliza NADH para reducir la FAD, esencial para la triptófano halogenasa BorH en la biosíntesis de la borregomicina A. El análisis estructural revela que el FAD debe separarse completamente del BorH para la reducción del BorF.

Área de la Ciencia:

La bioquímica
Biología estructural
Microbiología

Sus antecedentes:

BorF es una flavina reductasa de cadena corta de bacterias del suelo del desierto.
Utiliza NADH para reducir el dinucleótido de adenina flavina (FAD) a FADH.
Este FADH es crucial para la triptófano-6- halogenasa BorH en la biosíntesis de la borregomicina A.

Objetivo del estudio:

Para aclarar la base estructural de la reducción de FAD por BorF.
Para comprender la interacción entre BorF y FAD.
Comparar las estructuras BorF y BorH para inferir su relación funcional.

Principales métodos:

Cristalografía de rayos X para determinar la estructura del BorF unido al FAD a una resolución de 2,37 Å.
Técnica de sustitución molecular para la solución de estructura.
Análisis estructural comparativo del BorF y del BorH.

Principales resultados:

La estructura cristalina de rayos X de BorF reveló un homodímero con una topología de barril β dividido de clave griega.
La estructura muestra una hélice α N-terminal con intercambio de dominio, característica de esta familia de enzimas.
Las variaciones en las regiones del bucle entre α3 y β5 influyen en las conformaciones de unión de FAD.
La comparación estructural sugiere que la disociación de FAD de BorH es necesaria para la reducción mediada por BorF.

Conclusiones:

BorF posee un pliegue estructural conservado, pero exhibe variaciones únicas en el sitio de unión de FAD.
El estudio proporciona información sobre el mecanismo de reducción de la FAD por las flavinas reductasas de cadena corta.
Se propone la disociación completa de FAD del BorH como requisito previo para su reducción por BorF, destacando la dinámica enzima-sustrato.

Palabras clave:

Dinucleótido de adenina flavina Biosíntesis La cristalografía La oxidorreductaza

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