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El ensamblaje biomimético y la activación de [FeFe]-hidrogenases
G Berggren1,2, A Adamska3, C Lambertz4
1Laboratoire de Chimie et Biologie des Métaux (CEA / Université Grenoble 1 / CNRS), 17 rue des Martyrs, F-38054 Grenoble cedex 9, France.
Nature
|June 28, 2013
Resumen
Los investigadores activaron las [FeFe]-hidrogenasas utilizando imitaciones sintéticas. Un puente de azaditiolato específico activó con éxito la enzima, proporcionando información sobre la maduración de la hidrogenasa y permitiendo la producción de enzimas recombinantes.
Área de la Ciencia:
- La bioquímica es la bioquímica.
- Química bioorgánica Química bioorgánica.
- Enzimología Enzimología.
Sus antecedentes:
- Las hidrogenasas son catalizadores clave para la producción y absorción de hidrógeno, cruciales para la tecnología de celdas de combustible.
- El sitio activo de las [FeFe]-hidrogenasas contiene un centro único de di-hierro (subsitio [2Fe]) ensamblado a través de una vía biosintética compleja que involucra a la enzima de maduración HydF.
- Se han creado imitaciones sintéticas del subsitio [2Fe] pero carecen de actividad enzimática natural.
Objetivo del estudio:
- Para investigar la activación controlada de las [FeFe]-hidrogenasas utilizando imitaciones sintéticas del sitio activo.
- Para determinar el papel del ligando puente en el subsitio [2Fe] para la activación de la enzima.
- Desarrollar un método simplificado para la producción de hidrogenasas recombinantes activas.
Principales métodos:
- Se prepararon tres imitaciones sintéticas con diferentes ligandos de ditiolatos.
- Estos imitadores fueron cargados en la enzima bacteriana de maduración Thermotoga maritima HydF.
- El HydF cargado fue utilizado para activar apo-HydA1, una hidrogenasa de Chlamydomonas reinhardtii.
Principales resultados:
- Se logró una carga y transferencia exitosas de imitaciones sintéticas a apo-HydA1.
- La activación completa de HydA1 sólo se observó con el híbrido HydF que contiene el azadithiolate que imita el puente.
- Esto confirma la presencia de un ligando azaditiolato en las hidrogenasas [FeFe] nativas.
Conclusiones:
- La activación controlada de la metalloenzima es posible utilizando andamios de proteínas específicos y análogos sintéticos del sitio activo.
- El puente azaditiolato es esencial para la actividad de las [FeFe]-hidrogenasas nativas.
- Esta metodología ofrece conocimientos mecanicistas y estructurales sobre la maduración de la hidrogenasa y una herramienta para producir hidrogenasas recombinantes sin una maquinaria nativa completa.
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