Insights into the role of nucleotide-dependent conformational change in nitrogenase catalysis: Structural characterization of the nitrogenase Fe protein Leu127 deletion variant with bound MgATP

Sen, Sanchayita; Krishnakumar, Arathi; McClead, Jammi; Johnson, Michael K.; Seefeldt, Lance C.; Szilágyi, Róbert K.; Peters, John W.

doi:10.1016/j.jinorgbio.2006.02.016

Cited by 22 publications

(13 citation statements)

References 55 publications

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“…Additionally, a single residue deletion variant (∆L127-NifH) that is incapable of nucleotide hydrolysis was also crystallized with MgATP bound, and this structure is similarly unchanged compared to the MgADP-bound and nucleotide free structures [47]. However, as solid-state snapshots, these structures are not necessarily representative of the conformational dynamics operative in solution.…”

Section: Nucleotide Binding To Nifhmentioning

confidence: 99%

“…When the crystal structure of the Fe protein from A. vinelandii was solved in 1992 [2], a Walker's motif A [51] was identified between residues 9 and 16 that was indicative of a nucleotide binding site [52]. The partial occupancy of an ADP molecule in the 1992 crystal structure gave preliminary indications of how the nucleotide could bind to NifH, but a series of subsequent structures were solved with better-resolved MgADP [46] and MgATP [47] molecules.…”

Section: Nucleotide Binding To Nifhmentioning

confidence: 99%

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The Fe Protein: An Unsung Hero of Nitrogenase

Jasniewski

Sickerman

et al. 2018

Inorganics

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Although the nitrogen-fixing enzyme nitrogenase critically requires both a reductase component (Fe protein) and a catalytic component, considerably more work has focused on the latter species. Properties of the catalytic component, which contains two highly complex metallocofactors and catalyzes the reduction of N 2 into ammonia, understandably making it the "star" of nitrogenase. However, as its obligate redox partner, the Fe protein is a workhorse with multiple supporting roles in both cofactor maturation and catalysis. In particular, the nitrogenase Fe protein utilizes nucleotide binding and hydrolysis in concert with electron transfer to accomplish several tasks of critical importance. Aside from the ATP-coupled transfer of electrons to the catalytic component during substrate reduction, the Fe protein also functions in a maturase and insertase capacity to facilitate the biosynthesis of the two-catalytic component metallocofactors: fusion of the [Fe 8 S 7 ] P-cluster and insertion of Mo and homocitrate to form the matured [(homocitrate)MoFe 7 S 9 C] M-cluster. These and key structural-functional relationships of the indispensable Fe protein and its complex with the catalytic component will be covered in this review.

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Section: Nucleotide Binding To Nifhmentioning

confidence: 99%

Section: Nucleotide Binding To Nifhmentioning

confidence: 99%

The Fe Protein: An Unsung Hero of Nitrogenase

Jasniewski

Sickerman

et al. 2018

Inorganics

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“…The enzyme is composed of two-component metalloproteins called the iron (Fe) protein (also called dinitrogenase reductase or component II) and the molybdenum-iron (MoFe) protein (also called dinitrogenase or component I) ( Figure 1). The structures of the two nitrogenase component proteins (individually and in complex with each other), have been determined by X-ray crystallography (6)(7)(8)(9)(10)(11)(12)(13)(14)(15)(16)(17)(18)(19)(20)(21)(22)(23)(24)(25). These structures have provided detailed models of the three metal clusters contained in nitrogenase.…”

Section: Introductionmentioning

confidence: 99%

Mechanism of Mo-Dependent Nitrogenase

Yang

Danyal

Seefeldt

2011

Methods in Molecular Biology

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Nitrogenase is the enzyme responsible for biological reduction of dinitrogen (N(2)) to ammonia, a form usable for life. Playing a central role in the global biogeochemical nitrogen cycle, this enzyme has been the focus of intensive research for over 60 years. This chapter provides an overview of the features of nitrogenase as a background to the subsequent chapters of this volume that detail the many methods that have been applied in an attempt to gain a deeper understanding of this complex enzyme.

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“…The Fe protein is a homodimeric protein containing an [Fe 4 -S 4 ] cluster and two MgATP-binding sites [5]. The two halves of the Fe protein are flexible and adopt different conformations during ATP binding, complex formation and ATP hydrolysis [7]. The conformational change that occurs after association of the Fe protein with MgATP affects both the redox potential of the [Fe 4 -S 4 ] cluster and increases the affinity of the Fe protein for the MoFe protein.…”

mentioning

confidence: 99%

Electron transfer and half-reactivity in nitrogenase

Clarke

Fairhurst

Lowe

et al. 2011

Biochemical Society Transactions

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Nitrogenase is a globally important enzyme that catalyses the reduction of atmospheric dinitrogen into ammonia and is thus an important part of the nitrogen cycle. The nitrogenase enzyme is composed of a catalytic molybdenum-iron protein (MoFe protein) and a protein containing an [Fe4-S4] cluster (Fe protein) that functions as a dedicated ATP-dependent reductase. The current understanding of electron transfer between these two proteins is based on stopped-flow spectrophotometry, which has allowed the rates of complex formation and electron transfer to be accurately determined. Surprisingly, a total of four Fe protein molecules are required to saturate one MoFe protein molecule, despite there being only two well-characterized Fe-protein-binding sites. This has led to the conclusion that the purified Fe protein is only half-active with respect to electron transfer to the MoFe protein. Studies on the electron transfer between both proteins using rapid-quench EPR confirmed that, during pre-steady-state electron transfer, the Fe protein only becomes half-oxidized. However, stopped-flow spectrophotometry on MoFe protein that had only one active site occupied was saturated by approximately three Fe protein equivalents. These results imply that the Fe protein has a second interaction during the initial stages of mixing that is not involved in electron transfer.

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Insights into the role of nucleotide-dependent conformational change in nitrogenase catalysis: Structural characterization of the nitrogenase Fe protein Leu127 deletion variant with bound MgATP

Cited by 22 publications

References 55 publications

The Fe Protein: An Unsung Hero of Nitrogenase

The Fe Protein: An Unsung Hero of Nitrogenase

Mechanism of Mo-Dependent Nitrogenase

Electron transfer and half-reactivity in nitrogenase

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