Azotobacter vinelandii glutaredoxin D delivers the core cluster to nitrogenase cofactor scaffold protein NifU

Collantes, J. A.; Rosa-Nunez, E.; Armas, A. M.; Raimunda, D. C.; Perez-Gonzalez, A.; Guo, Y.; Echavarri-Erasun, C.; Rubio, L. M.; Gonzalez-Guerrero, M.

doi:10.1101/2025.10.21.683637

bioRxiv · 10.1101/2025.10.21.683637

Azotobacter vinelandii glutaredoxin D delivers the core cluster to nitrogenase cofactor scaffold protein NifU

Collantes, J. A.·Rosa-Nunez, E.·Armas, A. M.·Raimunda, D. C.·Perez-Gonzalez, A.·Guo, Y.·Echavarri-Erasun, C.·Rubio, L. M.·Gonzalez-Guerrero, M.

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Abstract

The scaffold protein NifU plays a central role in assembling the precursor [Fe4S4] clusters required for nitrogenase to function. The synthesis of these precursors depends on a catalytic [Fe2S2] group within NifU core ferredoxin domain. Here, we show that the monothiol glutaredoxin GrxD delivers this cluster to the NifU scaffold protein. Consistently, grxD mutants have reduced nitrogenase activity, the result of altered iron allocation to this enzyme. Biochemical assays show that GrxD unidirectionally transfers [Fe2S2] to NifU through protein-protein interaction. This allows GrxD to restore apo-NifU functionality, enabling proper [Fe4S4] synthesis, and NifH activation. These findings are crucial to understand how iron is allocated to nitrogenase for biological nitrogen fixation.

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Collantes, J. A., Rosa-Nunez, E., Armas, A. M., Raimunda, D. C., Perez-Gonzalez, A., Guo, Y., Echavarri-Erasun, C., Rubio, L. M., Gonzalez-Guerrero, M.. 2025-10-21. Azotobacter vinelandii glutaredoxin D delivers the core cluster to nitrogenase cofactor scaffold protein NifU. https://doi.org/10.1101/2025.10.21.683637

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