Fe protein-independent substrate reduction by nitrogenase MoFe protein variants

Karamatullah Danyal, Andrew J. Rasmussen, Stephen M. Keable, Boyd S. Inglet, Sudipta Shaw, Oleg A. Zadvornyy, Simon Duval, Dennis R. Dean, Simone Raugei, John W. Peters, Lance C. Seefeldt

Research output: Contribution to journalArticle

12 Citations (Scopus)

Abstract

The reduction of substrates catalyzed by nitrogenase normally requires nucleotide-dependent Fe protein delivery of electrons to the MoFe protein, which contains the active site FeMo cofactor. Here, it is reported that independent substitution of three amino acids (β-98Tyr→His, α-64Tyr→His, and β-99Phe→His) located between the P cluster and FeMo cofactor within the MoFe protein endows it with the ability to reduce protons to H2, azide to ammonia, and hydrazine to ammonia without the need for Fe protein or ATP. Instead, electrons can be provided by the low-potential reductant polyaminocarboxylate-ligated Eu(II) (Em values of -1.1 to -0.84 V vs the normal hydrogen electrode). The crystal structure of the β-98Tyr→His variant MoFe protein was determined, revealing only small changes near the amino acid substitution that affect the solvent structure and the immediate vicinity between the P cluster and the FeMo cofactor, with no global conformational changes observed. Computational normal-mode analysis of the nitrogenase complex reveals coupling in the motions of the Fe protein and the region of the MoFe protein with these three amino acids, which suggests a possible mechanism for how Fe protein might communicate subtle changes deep within the MoFe protein that profoundly affect intramolecular electron transfer and substrate reduction.

Original languageEnglish
Pages (from-to)2456-2462
Number of pages7
JournalBiochemistry
Volume54
Issue number15
DOIs
Publication statusPublished - Apr 21 2015

Fingerprint

Molybdoferredoxin
Nitrogenase
Substrates
Proteins
hydrazine
Electrons
Amino Acid Substitution
Ammonia
Amino Acids
Substitution reactions
Azides
Reducing Agents
Protons
Hydrogen
Catalytic Domain
Electrodes
Nucleotides
Adenosine Triphosphate
Crystal structure

ASJC Scopus subject areas

  • Biochemistry
  • Medicine(all)

Cite this

Danyal, K., Rasmussen, A. J., Keable, S. M., Inglet, B. S., Shaw, S., Zadvornyy, O. A., ... Seefeldt, L. C. (2015). Fe protein-independent substrate reduction by nitrogenase MoFe protein variants. Biochemistry, 54(15), 2456-2462. https://doi.org/10.1021/acs.biochem.5b00140

Fe protein-independent substrate reduction by nitrogenase MoFe protein variants. / Danyal, Karamatullah; Rasmussen, Andrew J.; Keable, Stephen M.; Inglet, Boyd S.; Shaw, Sudipta; Zadvornyy, Oleg A.; Duval, Simon; Dean, Dennis R.; Raugei, Simone; Peters, John W.; Seefeldt, Lance C.

In: Biochemistry, Vol. 54, No. 15, 21.04.2015, p. 2456-2462.

Research output: Contribution to journalArticle

Danyal, K, Rasmussen, AJ, Keable, SM, Inglet, BS, Shaw, S, Zadvornyy, OA, Duval, S, Dean, DR, Raugei, S, Peters, JW & Seefeldt, LC 2015, 'Fe protein-independent substrate reduction by nitrogenase MoFe protein variants', Biochemistry, vol. 54, no. 15, pp. 2456-2462. https://doi.org/10.1021/acs.biochem.5b00140
Danyal K, Rasmussen AJ, Keable SM, Inglet BS, Shaw S, Zadvornyy OA et al. Fe protein-independent substrate reduction by nitrogenase MoFe protein variants. Biochemistry. 2015 Apr 21;54(15):2456-2462. https://doi.org/10.1021/acs.biochem.5b00140
Danyal, Karamatullah ; Rasmussen, Andrew J. ; Keable, Stephen M. ; Inglet, Boyd S. ; Shaw, Sudipta ; Zadvornyy, Oleg A. ; Duval, Simon ; Dean, Dennis R. ; Raugei, Simone ; Peters, John W. ; Seefeldt, Lance C. / Fe protein-independent substrate reduction by nitrogenase MoFe protein variants. In: Biochemistry. 2015 ; Vol. 54, No. 15. pp. 2456-2462.
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