The Fe-only nitrogenase and the Mo nitrogenase from Rhodobacter capsulatus - A comparative study on the redox properties of the metal clusters present in the dinitrogenase components

Siemann S, Schneider K, Drottboom M, Müller A (2002)
EUROPEAN JOURNAL OF BIOCHEMISTRY 269(6): 1650-1661.

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Zeitschriftenaufsatz | Veröffentlicht | Englisch
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Abstract / Bemerkung
The dinitrogenase component proteins of the conventional Mo nitrogenase (MoFe protein) and of the alternative Fe-only nitrogenase (FeFe protein) were both isolated and purified from Rhodobacter capsulatus, redox-titrated according to the same procedures and subjected to an EPR spectroscopic comparison. In the course of an oxidative titration of the MoFe protein (Rc1(Mo)) three significant S = 1/2 EPR signals deriving from oxidized states of the P-cluster were detected: (1) a rhombic signal (g = 2.07, 1.96 and 1.83), which showed a bell-shaped redox curve with midpoint potentials (E-m) of -195 mV (appearance) and -30 mV (disappearance), (2) an axial signal (g(ii) = 2.00, g(perpendicular to) = 1.90) with almost identical redox properties and (3) a second rhombic signal (g = 2.03, 2.00, 1.90) at higher redox potentials ( > 100 mV). While the 'low-potential' rhombic signal and the axial signal have been both attributed to the one-electron-oxidized P-cluster (P1+) present in two conformationally different proteins, the 'high-potential' rhombic signal has been suggested rather to derive from the P3+ state. Upon oxidation, the FeFe protein (Rc1(Fe)) exibited three significant S = 1/2 EPR signals as well. However, the Re I F, signals strongly deviated from the MoFe protein signals, suggesting that they cannot simply be assigned to different P-cluster states. (a) The most prominent feature is an unusually broad signal at g = 2.27 and 2.06, which proved to be fully reversible and to correlate with catalytic activity. The cluster giving rise to this signal appears to be involved in the transfer of two electrons. The midpoint potentials determined were: -80 mV (appearance) and 70 mV (disappearance). (b) Under weakly acidic conditions (pH 6.4) a slightly altered EPR signal occurred. It was characterized by a shift of the g values to 2.22 and 2.05 and by the appearance of an additional negative absorption-shaped peak at g = 1.86. (c) A very narrow rhombic EPR signal at g = 2.00, 1.98 and 1.96 appeared at positive redox potentials (E-m = 80 mV, intensity maximum at 160 mV). Another novel S = 1/2 signal at g = 1.96, 1.92 and 1.77 was observed on further, enzymatic reduction of the dithionite-reduced state of Rc1(Fe), with the dinitrogenase reductase component (Rc2(Fe)) of the same enzyme system (turnover conditions in the presence of N-2 and ATP). When the Rc1(Mo) protein was treated analogously. neither this 'turnover signal' nor any other S = 1/2 signal were detectable. All Rc1(Fe)-specific EPR signals detected are discussed and tentatively assigned with special consideration of the reference spectra obtained from Rc1(Mo) preparations.
Erscheinungsjahr
Zeitschriftentitel
EUROPEAN JOURNAL OF BIOCHEMISTRY
Band
269
Ausgabe
6
Seite(n)
1650-1661
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Siemann S, Schneider K, Drottboom M, Müller A. The Fe-only nitrogenase and the Mo nitrogenase from Rhodobacter capsulatus - A comparative study on the redox properties of the metal clusters present in the dinitrogenase components. EUROPEAN JOURNAL OF BIOCHEMISTRY. 2002;269(6):1650-1661.
Siemann, S., Schneider, K., Drottboom, M., & Müller, A. (2002). The Fe-only nitrogenase and the Mo nitrogenase from Rhodobacter capsulatus - A comparative study on the redox properties of the metal clusters present in the dinitrogenase components. EUROPEAN JOURNAL OF BIOCHEMISTRY, 269(6), 1650-1661. doi:10.1046/j.1432-1327.2002.02804.x
Siemann, S., Schneider, K., Drottboom, M., and Müller, A. (2002). The Fe-only nitrogenase and the Mo nitrogenase from Rhodobacter capsulatus - A comparative study on the redox properties of the metal clusters present in the dinitrogenase components. EUROPEAN JOURNAL OF BIOCHEMISTRY 269, 1650-1661.
Siemann, S., et al., 2002. The Fe-only nitrogenase and the Mo nitrogenase from Rhodobacter capsulatus - A comparative study on the redox properties of the metal clusters present in the dinitrogenase components. EUROPEAN JOURNAL OF BIOCHEMISTRY, 269(6), p 1650-1661.
S. Siemann, et al., “The Fe-only nitrogenase and the Mo nitrogenase from Rhodobacter capsulatus - A comparative study on the redox properties of the metal clusters present in the dinitrogenase components”, EUROPEAN JOURNAL OF BIOCHEMISTRY, vol. 269, 2002, pp. 1650-1661.
Siemann, S., Schneider, K., Drottboom, M., Müller, A.: The Fe-only nitrogenase and the Mo nitrogenase from Rhodobacter capsulatus - A comparative study on the redox properties of the metal clusters present in the dinitrogenase components. EUROPEAN JOURNAL OF BIOCHEMISTRY. 269, 1650-1661 (2002).
Siemann, S, Schneider, Klaus, Drottboom, M, and Müller, Achim. “The Fe-only nitrogenase and the Mo nitrogenase from Rhodobacter capsulatus - A comparative study on the redox properties of the metal clusters present in the dinitrogenase components”. EUROPEAN JOURNAL OF BIOCHEMISTRY 269.6 (2002): 1650-1661.

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Nitrogenase and homologs.
Hu Y, Ribbe MW., J Biol Inorg Chem 20(2), 2015
PMID: 25491285

51 References

Daten bereitgestellt von Europe PubMed Central.

Isolation of a new vanadium-containing nitrogenase from Azotobacter vinelandii.
Hales BJ, Case EE, Morningstar JE, Dzeda MF, Mauterer LA., Biochemistry 25(23), 1986
PMID: 3026449
The vanadium nitrogenase of Azotobacter chroococcum. Purification and properties of the VFe protein.
Eady RR, Robson RL, Richardson TH, Miller RW, Hawkins M., Biochem. J. 244(1), 1987
PMID: 2821997
Structure, function, and biosynthesis of the metallosulfur clusters in nitrogenases
Smith, Adv. Inorg. Chem. 47(), 1999
Purification of a second alternative nitrogenase from a nifHDK deletion strain of Azotobacter vinelandii.
Chisnell JR, Premakumar R, Bishop PE., J. Bacteriol. 170(1), 1988
PMID: 3121587
Comparative biochemical characterization of the iron-only nitrogenase and the molybdenum nitrogenase from Rhodobacter capsulatus.
Schneider K, Gollan U, Drottboom M, Selsemeier-Voigt S, Muller A., Eur. J. Biochem. 244(3), 1997
PMID: 9108249
Purification and characterization of the alternative nitrogenase from the photosynthetic bacterium
Davis, Rhodospirillum rubrum. J. Bacteriol. 178(), 1996
Redox-dependent structural changes in the nitrogenase P-cluster.
Peters JW, Stowell MH, Soltis SM, Finnegan MG, Johnson MK, Rees DC., Biochemistry 36(6), 1997
PMID: 9063865
Nitrogenase: a nucleotide-dependent molecular switch.
Howard JB, Rees DC., Annu. Rev. Biochem. 63(), 1994
PMID: 7979238
How N2 might be activated by the FeMo-cofactor in nitrogenase
Deng, Angew. Chem. Int. Ed. 32(), 1993
Elementary reactions, structure-function relationships, and the potential relevance of low molecular weight metal-sulfur ligand complexes to biological N2 fixation
Sellmann, J. Biol. Inorg. Chem. 1(), 1997
Functional analogs for the reduction of certain nitrogenase substrates. Are multiple sites within the Fe/Mo/S center involved in the 6e- reduction of N2?
Coucouvanis, J. Biol. Inorg. Chem. 1(), 1996
Why R-homocitrate is essential to the reactivity of FeMo-cofactor of nitrogenase. Studies on NifV- extracted FeMo-cofactor
Grönberg, J. Am. Chem. Soc. 120(), 1998

Krahn, 1998
The Fe-only nitrogenase from Rhodobacter capsulatus: identification of the cofactor, an unusual, high-nuclearity iron-sulfur cluster, by Fe K-edge EXAFS and 57Fe Mossbauer spectroscopy.
Krahn E, Weiss R, Krockel M, Groppe J, Henkel G, Cramer P, Trautwein X, Schneider K, Muller A., J. Biol. Inorg. Chem. 7(1-2), 2001
PMID: 11862539
Diphosphofructose- aldolase, Phosphoglyceraldehyd-dehydrogenase, Milchsäure-dehydrogenase, Glycerophosphat-dehydrogenase and Pyruvat-kinase aus Kaninchenmuskulatur in einem Arbeitsgang
Beisenherz, Z. Naturforsch. 8b(), 1953
FeMo cofactor biosynthesis in a nifE- mutant of Rhodobacter capsulatus.
Siemann S, Schneider K, Behrens K, Knochel A, Klipp W, Muller A., Eur. J. Biochem. 268(7), 2001
PMID: 11277916
Redox properties and EPR spectroscopy of the P clusters of Azotobacter vinelandii MoFe protein.
Pierik AJ, Wassink H, Haaker H, Hagen WR., Eur. J. Biochem. 212(1), 1993
PMID: 8383042
Detection of EPR signals assigned to the 1-equivalent-oxidized P-clusters of the nitrogenase MoFe protein from Azotobacter vinelandii
Tittsworth, J. Am. Chem. Soc. 115(), 1993
Evidence for coupled electron and proton transfer in the [8Fe-7S] cluster of nitrogenase.
Lanzilotta WN, Christiansen J, Dean DR, Seefeldt LC., Biochemistry 37(32), 1998
PMID: 9698385
Spectroscopic evidence for changes in the redox state of the nitrogenase P-cluster during turnover.
Chan JM, Christiansen J, Dean DR, Seefeldt LC., Biochemistry 38(18), 1999
PMID: 10231529
Mössbauer study of the MoFe protein of nitrogenase from Azotobacter vinelandii using selective 57Fe enrichment of the M centers
Yoo, J. Am. Chem. Soc. 122(), 2000

Hagen, 1989
The molybdenum nitrogenase from wild-type Xanthobacter autotrophicus exhibits properties reminiscent of alternative nitrogenases.
Schneider K, Muller A, Krahn E, Hagen WR, Wassink H, Knuttel KH., Eur. J. Biochem. 230(2), 1995
PMID: 7607241
Isolated iron-molybdenum cofactor of nitrogenase exists in multiple forms in its oxidized and semi-reduced states.
Newton WE, Gheller SF, Feldman BJ, Dunham WR, Schultz FA., J. Biol. Chem. 264(4), 1989
PMID: 2536693
The electron paramagnetic resonance of metalloproteins.
Palmer G., Biochem. Soc. Trans. 13(3), 1985
PMID: 2993061
EPR and Mössbauer spectroscopic studies on the tetrameric, NAD-linked hydrogenase of Nocardia opaca 1b and its two dimers: 1. The βδ-dimer - a prototype of a simple hydrogenase
Zaborosch, Biometals 8(), 1995
The iron-molybdenum cofactor of nitrogenase
Burgess, Chem. Rev. 90(), 1990
Mössbauer and integer-spin EPR of the oxidized P-clusters of nitrogenase: Pox is a non-Kramers system with a nearly degenerate ground doublet
Surerus, J. Am. Chem. Soc. 114(), 1992

Orme-Johnson, 1993
The electronic structure of FeS centres in proteins and models. A contribution to the understanding of their electron transfer properties
Bertini, Struct. Bonding 83(), 1995

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