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UvrC Coordina un cofactor [4Fe4S] O2-sensible

Rebekah M B Silva1, Michael A Grodick1, Jacqueline K Barton1

  • 1Division of Chemistry and Chemical Engineering, California Institute of Technology, Pasadena, California 91125, United States.

Journal of the American Chemical Society
|May 30, 2020
PubMed
Resumen
Este resumen es generado por máquina.

Los investigadores investigaron el grupo [4Fe4S] en UvrC, una enzima de reparación del ADN. Descubrieron que este cúmulo de hierro y azufre es crucial para la UvrC.

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Área de la Ciencia:

  • La bioquímica
  • Biología molecular
  • Química bioorgánica

Sus antecedentes:

  • Los grupos [4Fe4S] son cofactores vitales en las enzimas en todos los dominios de la vida.
  • La observación de estos grupos en las enzimas de procesamiento de ácido nucleico es difícil debido a las firmas débiles y la labilidad del cofactor.
  • UvrC, una endonucleasa de reparación por escisión de nucleótidos (NER) bacteriana, fue investigada para la participación de clúster [4Fe4S].

Objetivo del estudio:

  • Investigar la presencia y el papel de un grupo [4Fe4S] en la enzima UvrC.
  • Caracterizar la estabilidad del cofactor y su impacto en la afinidad y función de unión al ADN de UvrC.
  • Para determinar la actividad redox del grupo [4Fe4S] dentro de UvrC.

Principales métodos:

  • Alineaciones de secuencias y técnicas de purificación anaeróbica.
  • Espectroscopias de cuantificación de hierro, visibles por rayos UV y de resonancia paramagnética electrónica (EPR).
  • Pruebas de unión al ADN y electroquímica del ADN.

Principales resultados:

  • El UvrC exhibe características consistentes con la coordinación del clúster [4Fe4S] (incorporación del 60-70%).
  • El grupo [4Fe4S] en UvrC es susceptible a la degradación oxidativa.
  • Holo-UvrC muestra una alta afinidad de unión al ADN (80-100 nM) en comparación con apo-UvrC.
  • El grupo [4Fe4S] es redox-activo y participa en el transporte de carga mediado por ADN (potencial de punto medio de 90 mV vs NHE).

Conclusiones:

  • El clúster [4Fe4S] es crítico para la función de UvrC, mejorando su afinidad de unión al ADN.
  • La actividad redox del cofactor sugiere un papel en los mecanismos de reparación del ADN más allá de la unión simple.
  • Comprender el clúster [4Fe4S] en UvrC proporciona información sobre las vías de reparación del ADN y la función de las metalloenzimas.