Structural characterization of the P1 intermediate state of the P-cluster of nitrogenase

Stephen M. Keable, Oleg A. Zadvornyy, Lewis E. Johnson, Bojana Ginovska, Andrew J. Rasmussen, Karamatullah Danyal, Brian J. Eilers, Gregory A. Prussia, Axl X. LeVan, Simone Raugei, Lance C. Seefeldt, John W. Peters

Research output: Contribution to journalArticle

Abstract

Nitrogenase is the enzyme that reduces atmospheric dinitrogen (N2) to ammonia (NH3) in biological systems. It catalyzes a series of single-electron transfers from the donor iron protein (Fe protein) to the molybdenum–iron protein (MoFe protein) that contains the iron–molybdenum cofactor (FeMo-co) sites where N2 is reduced to NH3. The P-cluster in the MoFe protein functions in nitrogenase catalysis as an intermediate electron carrier between the external electron donor, the Fe protein, and the FeMo-co sites of the MoFe protein. Previous work has revealed that the P-cluster undergoes redox-dependent structural changes and that the transition from the all-ferrous resting (PN) state to the two-electron oxidized P2 state is accompanied by protein serine hydroxyl and backbone amide ligation to iron. In this work, the MoFe protein was poised at defined potentials with redox mediators in an electrochemical cell, and the three distinct structural states of the P-cluster (P2, P1, and PN) were characterized by X-ray crystallography and confirmed by computational analysis. These analyses revealed that the three oxidation states differ in coordination, implicating that the P1 state retains the serine hydroxyl coordination but lacks the backbone amide coordination observed in the P2 states. These results provide a complete picture of the redox-dependent ligand rearrangements of the three P-cluster redox states.

Original languageEnglish
Pages (from-to)9629-9635
Number of pages7
JournalJournal of Biological Chemistry
Volume293
Issue number25
DOIs
Publication statusPublished - Jan 1 2018

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Molybdoferredoxin
Nitrogenase
Oxidation-Reduction
Electrons
Proteins
Amides
Hydroxyl Radical
Serine
Iron
Electrochemical cells
X ray crystallography
X Ray Crystallography
Biological systems
Ataxia
Catalysis
Ammonia
Ligation
Ligands
Oxidation
Enzymes

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology
  • Cell Biology

Cite this

Keable, S. M., Zadvornyy, O. A., Johnson, L. E., Ginovska, B., Rasmussen, A. J., Danyal, K., ... Peters, J. W. (2018). Structural characterization of the P1 intermediate state of the P-cluster of nitrogenase. Journal of Biological Chemistry, 293(25), 9629-9635. https://doi.org/10.1074/jbc.RA118.002435

Structural characterization of the P1 intermediate state of the P-cluster of nitrogenase. / Keable, Stephen M.; Zadvornyy, Oleg A.; Johnson, Lewis E.; Ginovska, Bojana; Rasmussen, Andrew J.; Danyal, Karamatullah; Eilers, Brian J.; Prussia, Gregory A.; LeVan, Axl X.; Raugei, Simone; Seefeldt, Lance C.; Peters, John W.

In: Journal of Biological Chemistry, Vol. 293, No. 25, 01.01.2018, p. 9629-9635.

Research output: Contribution to journalArticle

Keable, SM, Zadvornyy, OA, Johnson, LE, Ginovska, B, Rasmussen, AJ, Danyal, K, Eilers, BJ, Prussia, GA, LeVan, AX, Raugei, S, Seefeldt, LC & Peters, JW 2018, 'Structural characterization of the P1 intermediate state of the P-cluster of nitrogenase', Journal of Biological Chemistry, vol. 293, no. 25, pp. 9629-9635. https://doi.org/10.1074/jbc.RA118.002435
Keable SM, Zadvornyy OA, Johnson LE, Ginovska B, Rasmussen AJ, Danyal K et al. Structural characterization of the P1 intermediate state of the P-cluster of nitrogenase. Journal of Biological Chemistry. 2018 Jan 1;293(25):9629-9635. https://doi.org/10.1074/jbc.RA118.002435
Keable, Stephen M. ; Zadvornyy, Oleg A. ; Johnson, Lewis E. ; Ginovska, Bojana ; Rasmussen, Andrew J. ; Danyal, Karamatullah ; Eilers, Brian J. ; Prussia, Gregory A. ; LeVan, Axl X. ; Raugei, Simone ; Seefeldt, Lance C. ; Peters, John W. / Structural characterization of the P1 intermediate state of the P-cluster of nitrogenase. In: Journal of Biological Chemistry. 2018 ; Vol. 293, No. 25. pp. 9629-9635.
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