Identification of two tetranuclear FeS clusters on the ferredoxin-type subunit of NADH : ubiquinone oxidoreductase (complex I)

Tim Rasmussen, D Scheide, B Brors, L Kintscher, H Weiss, T Friedrich

Research output: Contribution to journalArticle

65 Citations (Scopus)

Abstract

The proton-translocating NADH:ubiquinone oxidoreductase of respiratory chains (complex I) contains one flavin mononucleotide and five EPR-detectable iron-sulfur clusters as redox groups. Because of the number of conserved motifs typical for binding iron-sulfur clusters and the high content of iron and acid-labile sulfide of complex I preparations, it is predicted that complex I contains additional clusters which have not yet been detected by EPR spectroscopy. To search for such clusters, we used a combination of UV/vis and EPR spectroscopy to study complex I from Neurospora crassa and Escherichia coli adjusted to distinct redox states. We detected a UV/vis redox difference spectrum characterized by negative absorbances at 325 and 425 nm that could not be assigned to the known redox groups. Redox titration was used to determine the pH-independent midpoint potential to be -270 mV, being associated with the transfer of two electrons. Comparison with UV/vis difference spectra obtained from complex I fragments and related enzymes showed that this group is localized on subunit Nuo21.3c of the N, crassa or NuoI of the E. coli complex I, respectively. This subunit (the bovine TYKY) belongs to a family of 8Fe-ferredoxins which contain two tetranuclear iron-sulfur clusters as redox groups. We detected EPR signals in a fragment of complex I which we attribute to the novel FeS clusters of complex I.

Original languageEnglish
Pages (from-to)6124-6131
Number of pages8
JournalBiochemistry
Volume40
Issue number20
DOIs
Publication statusPublished - 22 May 2001

Keywords

  • IRON-SULFUR CLUSTERS
  • MITOCHONDRIAL NADH
  • ESCHERICHIA-COLI
  • CLOSTRIDIUM-PASTEURIANUM
  • REDUCING HYDROGENASE
  • NEUROSPORA-CRASSA
  • ALCALIGENES-EUTROPHUS
  • DEHYDROGENASE
  • BACTERIAL
  • REDUCTION
  • IRON SULFUR CLUSTERS
  • ESCHERICHIA COLI
  • CLOSTRIDIUM PASTEURIANUM
  • NEUROSPORA CRASSA
  • ALCALIGENES EUTROPHUS
  • REDUCTION

Cite this

Identification of two tetranuclear FeS clusters on the ferredoxin-type subunit of NADH : ubiquinone oxidoreductase (complex I). / Rasmussen, Tim; Scheide, D ; Brors, B ; Kintscher, L ; Weiss, H ; Friedrich, T .

In: Biochemistry, Vol. 40, No. 20, 22.05.2001, p. 6124-6131.

Research output: Contribution to journalArticle

Rasmussen, T, Scheide, D, Brors, B, Kintscher, L, Weiss, H & Friedrich, T 2001, 'Identification of two tetranuclear FeS clusters on the ferredoxin-type subunit of NADH : ubiquinone oxidoreductase (complex I)', Biochemistry, vol. 40, no. 20, pp. 6124-6131. https://doi.org/10.1021/bi0026977
Rasmussen, Tim ; Scheide, D ; Brors, B ; Kintscher, L ; Weiss, H ; Friedrich, T . / Identification of two tetranuclear FeS clusters on the ferredoxin-type subunit of NADH : ubiquinone oxidoreductase (complex I). In: Biochemistry. 2001 ; Vol. 40, No. 20. pp. 6124-6131.
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abstract = "The proton-translocating NADH:ubiquinone oxidoreductase of respiratory chains (complex I) contains one flavin mononucleotide and five EPR-detectable iron-sulfur clusters as redox groups. Because of the number of conserved motifs typical for binding iron-sulfur clusters and the high content of iron and acid-labile sulfide of complex I preparations, it is predicted that complex I contains additional clusters which have not yet been detected by EPR spectroscopy. To search for such clusters, we used a combination of UV/vis and EPR spectroscopy to study complex I from Neurospora crassa and Escherichia coli adjusted to distinct redox states. We detected a UV/vis redox difference spectrum characterized by negative absorbances at 325 and 425 nm that could not be assigned to the known redox groups. Redox titration was used to determine the pH-independent midpoint potential to be -270 mV, being associated with the transfer of two electrons. Comparison with UV/vis difference spectra obtained from complex I fragments and related enzymes showed that this group is localized on subunit Nuo21.3c of the N, crassa or NuoI of the E. coli complex I, respectively. This subunit (the bovine TYKY) belongs to a family of 8Fe-ferredoxins which contain two tetranuclear iron-sulfur clusters as redox groups. We detected EPR signals in a fragment of complex I which we attribute to the novel FeS clusters of complex I.",
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T1 - Identification of two tetranuclear FeS clusters on the ferredoxin-type subunit of NADH

T2 - ubiquinone oxidoreductase (complex I)

AU - Rasmussen, Tim

AU - Scheide, D

AU - Brors, B

AU - Kintscher, L

AU - Weiss, H

AU - Friedrich, T

PY - 2001/5/22

Y1 - 2001/5/22

N2 - The proton-translocating NADH:ubiquinone oxidoreductase of respiratory chains (complex I) contains one flavin mononucleotide and five EPR-detectable iron-sulfur clusters as redox groups. Because of the number of conserved motifs typical for binding iron-sulfur clusters and the high content of iron and acid-labile sulfide of complex I preparations, it is predicted that complex I contains additional clusters which have not yet been detected by EPR spectroscopy. To search for such clusters, we used a combination of UV/vis and EPR spectroscopy to study complex I from Neurospora crassa and Escherichia coli adjusted to distinct redox states. We detected a UV/vis redox difference spectrum characterized by negative absorbances at 325 and 425 nm that could not be assigned to the known redox groups. Redox titration was used to determine the pH-independent midpoint potential to be -270 mV, being associated with the transfer of two electrons. Comparison with UV/vis difference spectra obtained from complex I fragments and related enzymes showed that this group is localized on subunit Nuo21.3c of the N, crassa or NuoI of the E. coli complex I, respectively. This subunit (the bovine TYKY) belongs to a family of 8Fe-ferredoxins which contain two tetranuclear iron-sulfur clusters as redox groups. We detected EPR signals in a fragment of complex I which we attribute to the novel FeS clusters of complex I.

AB - The proton-translocating NADH:ubiquinone oxidoreductase of respiratory chains (complex I) contains one flavin mononucleotide and five EPR-detectable iron-sulfur clusters as redox groups. Because of the number of conserved motifs typical for binding iron-sulfur clusters and the high content of iron and acid-labile sulfide of complex I preparations, it is predicted that complex I contains additional clusters which have not yet been detected by EPR spectroscopy. To search for such clusters, we used a combination of UV/vis and EPR spectroscopy to study complex I from Neurospora crassa and Escherichia coli adjusted to distinct redox states. We detected a UV/vis redox difference spectrum characterized by negative absorbances at 325 and 425 nm that could not be assigned to the known redox groups. Redox titration was used to determine the pH-independent midpoint potential to be -270 mV, being associated with the transfer of two electrons. Comparison with UV/vis difference spectra obtained from complex I fragments and related enzymes showed that this group is localized on subunit Nuo21.3c of the N, crassa or NuoI of the E. coli complex I, respectively. This subunit (the bovine TYKY) belongs to a family of 8Fe-ferredoxins which contain two tetranuclear iron-sulfur clusters as redox groups. We detected EPR signals in a fragment of complex I which we attribute to the novel FeS clusters of complex I.

KW - IRON-SULFUR CLUSTERS

KW - MITOCHONDRIAL NADH

KW - ESCHERICHIA-COLI

KW - CLOSTRIDIUM-PASTEURIANUM

KW - REDUCING HYDROGENASE

KW - NEUROSPORA-CRASSA

KW - ALCALIGENES-EUTROPHUS

KW - DEHYDROGENASE

KW - BACTERIAL

KW - REDUCTION

KW - IRON SULFUR CLUSTERS

KW - ESCHERICHIA COLI

KW - CLOSTRIDIUM PASTEURIANUM

KW - NEUROSPORA CRASSA

KW - ALCALIGENES EUTROPHUS

KW - REDUCTION

U2 - 10.1021/bi0026977

DO - 10.1021/bi0026977

M3 - Article

VL - 40

SP - 6124

EP - 6131

JO - Biochemistry

JF - Biochemistry

SN - 0006-2960

IS - 20

ER -