Participation of the disulfide bridge in the redox cycle of the ferredoxin from the hyperthermophile Pyrococcus furiosus: 1H nuclear magnetic resonance time resolution of the four redox states at ambient temperature

Cm, Gorst; Zh, Zhou; Ma, Kewei; Teng, Quincy; Jb, Howard; Adams, Michael W. W.; Gn, La Mar

doi:10.1021/bi00027a030

Cited by 36 publications

(56 citation statements)

References 25 publications

(32 reference statements)

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“…The differences in the NMR and Mo¨ssbauer spectra between FdII ox and FdII int were due to conformational changes resulting from the breaking/formation of the disulfide bridge. The same intermediate state was found later in the P. furiosus 4Fe Fd [67]. This may turn out to be a key observation for the mechanism of complex proteins containing iron-sulfur clusters.…”

Section: Introductionsupporting

confidence: 76%

Desulfovibrio gigas ferredoxin II: redox structural modulation of the [3Fe–4S] cluster

et al. 2006

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Desulfovibrio gigas ferredoxin II (DgFdII) is a small protein with a polypeptide chain composed of 58 amino acids, containing one Fe 3 S 4 cluster per monomer. Upon studying the redox cycle of this protein, we detected a stable intermediate (FdII int ) with four 1 H resonances at 24.1, 20.5, 20.8 and 13.7 ppm. The differences between FdII ox and FdII int were attributed to conformational changes resulting from the breaking/formation of an internal disulfide bridge. The same 1 H NMR methodology used to fully assign the three cysteinyl ligands of the [3Fe-4S] core in the oxidized state (DgFdII ox ) was used here for the assignment of the same three ligands in the intermediate state (DgFdII int ). The spin-coupling model used for the oxidized form of DgFdII where magnetic exchange coupling constants of around 300 cm À1 and hyperfine coupling constants equal to 1 MHz for all the three iron centres were found, does not explain the isotropic shift temperature dependence for the three cysteinyl cluster ligands in DgFdII int . This study, together with the spin delocalization mechanism proposed here for DgFdII int , allows the detection of structural modifications at the [3Fe-4S] cluster in DgFdII ox and DgFdII int .

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Section: Introductionsupporting

confidence: 76%

Desulfovibrio gigas ferredoxin II: redox structural modulation of the [3Fe–4S] cluster

et al. 2006

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“…A disulfide bond was detected in Pyrococcus furiosus ferrodoxin, in which it plays a role in the redox cycle of the protein (141). The crystal structures of DNA polymerases from Thermococcus gorgonarius and Thermococcus sp.…”

Section: Disulfide Bonds In Cytoplasmic Archaeal Proteinsmentioning

confidence: 99%

Posttranslational Protein Modification inArchaea

Eichler

Adams

2005

Microbiol Mol Biol Rev

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194

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One of the first hurdles to be negotiated in the postgenomic era involves the description of the entire protein content of the cell, the proteome. Such efforts are presently complicated by the various posttranslational modifications that proteins can experience, including glycosylation, lipid attachment, phosphorylation, methylation, disulfide bond formation, and proteolytic cleavage. Whereas these and other posttranslational protein modifications have been well characterized in Eucarya and Bacteria, posttranslational modification in Archaea has received far less attention. Although archaeal proteins can undergo posttranslational modifications reminiscent of what their eucaryal and bacterial counterparts experience, examination of archaeal posttranslational modification often reveals aspects not previously observed in the other two domains of life. In some cases, posttranslational modification allows a protein to survive the extreme conditions often encountered by Archaea. The various posttranslational modifications experienced by archaeal proteins, the molecular steps leading to these modifications, and the role played by posttranslational modification in Archaea form the focus of this review

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“…The positions of Cys 2~ (V) and Cys 43 (VI) in Tl Fd are homologous to the cysteines that participate in a disulfide bridge in the crystal structure of Dg 3Fe Fd (Kissinger et al, 1991) and in the solution structures of both 3Fe and 4Fe P f F d ~ (Teng et al, 1994;Gorst et al, 1995b). The present N M R data provide strong support for the presence of this disulfide bridge through strong and medium intensity NOESY cross peaks between the C~H and C~Hs of Cys 2~ (V) and the C~Hs of Cys 43 (V|).…”

Section: Disulfide Bridgementioning

confidence: 99%

“…TOCSY spectra (Bax and Davis, 1985) with slow repetition rates (0.6 s -1) and long mixing times (100 ms) were performed at three different temperatures (10, 30 and 40 ~ the WALTZ-16 pulse sequence (Shaka et al, 1983), with a soft pulse of 20-25 ~ts, was used as spin-lock. Standard NOESY experiments (Jeener et al, 1975) with long mixing time (300 ms) were performed at three different temperatures (10, 30 and 40 ~ A NOESY map with a mixing time of 50 ms and a more rapid repetition rate (2 s -l) was also collected at 40 ~ to detect dipolar connectivities among effectively relaxed signals (Gorst et al, 1995b). The data were collected over a spectral window of 7.017 kHz with 512 tl values, each consisting of 2048 t z points.…”

Section: Nmr Spectroscopymentioning

confidence: 99%

1H NMR investigation of the secondary structure, tertiary contacts and cluster environment of the four-iron ferredoxin from the hyperthermophilic archaeon Thermococcus litoralis

et al. 1996

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The solution molecular structure of the four-iron ferredoxin (Fd) from the hyperthermophilic archaeon Thermococcus litoralis (Tl) has been investigated by 1H NMR spectroscopy. TOCSY and NOESY experiments in H2O, tailored to detect both weakly and strongly relaxed resonances, together with steady-state NOEs in both H2O and D2O, allowed the identification of 58 of the 59 residues, with one residue near the paramagnetic center undetected. It is shown that the contact shifted and strongly relaxed signals for all four cysteines ligated to the paramagnetic cluster can be assigned by standard backbone connectivities that do not require any assumptions about the tertiary structure. Secondary structural elements identified in Tl Fd are a three-stranded antiparallel beta-strand involving the termini of the protein, a double beta-strand (also antiparallel), two alpha-helices and four turns. The existence of a disulfide bridge between the nonligated cysteines is also proposed. Dipolar contacts observed in the NOESY maps and by steady-state NOEs between the ligated cysteines and the 'diamagnetic' protein matrix indicate that the overall folding pattern of Tl Fd is very similar to that of the 3Fe ferredoxin from the mesophilic bacterium Desulfovibrio gigas [Kissinger et al. (1991) J. Mol. Biol., 219, 693-723]. The influence of the paramagnetism of the cluster on the relaxation properties of the proton signals of nonligated residues near the cluster, as well as on the ligated cysteines, correlates well with the proximity to the cluster iron(s), as predicted from the crystal structures for homologous protons of other single-cluster ferredoxins. Finally, the potential role of the various identified structural factors in contributing to the hyperthermostability of this protein is discussed.

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Participation of the disulfide bridge in the redox cycle of the ferredoxin from the hyperthermophile Pyrococcus furiosus: 1H nuclear magnetic resonance time resolution of the four redox states at ambient temperature

Cited by 36 publications

References 25 publications

Desulfovibrio gigas ferredoxin II: redox structural modulation of the [3Fe–4S] cluster

Desulfovibrio gigas ferredoxin II: redox structural modulation of the [3Fe–4S] cluster

Posttranslational Protein Modification inArchaea

1H NMR investigation of the secondary structure, tertiary contacts and cluster environment of the four-iron ferredoxin from the hyperthermophilic archaeon Thermococcus litoralis

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