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Literature summary for 7.2.1.2 extracted from

  • Backiel, J.; Juárez, O.; Zagorevski, D.; Wang, Z.; Nilges, M.; Barquera, B.
    Covalent binding of flavins to RnfG and RnfD in the Rnf complex from Vibrio cholerae (2008), Biochemistry, 47, 11273-11284.
    View publication on PubMedView publication on EuropePMC
Search for more literature for 7.2.1.2

Cloned(Commentary)

Cloned (Comment) Organism
genes rnfD and rnfG, cloning of Rnf complex subunits and recombinant individual expression of wild-type and mutant His-tagged RnfD and RnfG in Vibrio cholerae cells producing protein fluorescence under UV illumination, recombinant subunit protein topology analysis, overview Vibrio cholerae serotype O1

Protein Variants

Protein Variants Comment Organism
T175L site-directed mutagenesis of subunit RnfG, the mutation affects FMN cofactor binding Vibrio cholerae serotype O1
T187V site-directed mutagenesis of subunit RnfD, the mutation affects flavin cofactor binding Vibrio cholerae serotype O1
T278L site-directed mutagenesis of subunit RnfD, the mutation does not affect flavin cofactor binding Vibrio cholerae serotype O1

Localization

Localization Comment Organism GeneOntology No. Textmining
additional information the flavins in RnfG and RnfD are localized in the periplasmic space Vibrio cholerae serotype O1
-
-

Metals/Ions

Metals/Ions Comment Organism Structure
Fe2+ subunits RnfB and RnfC each contain the sequence motif for an iron-sulfur center Vibrio cholerae serotype O1

Molecular Weight [Da]

Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
26000
-
 1 * 26000, recombinant isolated subunit RnfG, SDS-PAGE, 1 * 39000, recombinant isolated subunit RnfD, SDS-PAGE Vibrio cholerae serotype O1
39000
-
 1 * 26000, recombinant isolated subunit RnfG, SDS-PAGE, 1 * 39000, recombinant isolated subunit RnfD, SDS-PAGE Vibrio cholerae serotype O1

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
2 reduced ferredoxin [iron-sulfur] cluster + NAD+ + H+ + Na+[side 1] Vibrio cholerae serotype O1
-
 2 oxidized ferredoxin [iron-sulfur] cluster + NADH + Na+[side 2]
-
 ?
2 reduced ferredoxin [iron-sulfur] cluster + NAD+ + H+ + Na+[side 1] Vibrio cholerae serotype O1 O395N-1
-
 2 oxidized ferredoxin [iron-sulfur] cluster + NADH + Na+[side 2]
-
 ?

Organism

Organism UniProt Comment Textmining
Vibrio cholerae serotype O1
-
-
-
Vibrio cholerae serotype O1 O395N-1
-
-
-

Purification (Commentary)

Purification (Comment) Organism
recombinant wild-type and mutant His-tagged RnfD and RnfG fom Vibrio cholerae cells by nickel affinity chromatography and gel filtration Vibrio cholerae serotype O1

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
2 reduced ferredoxin [iron-sulfur] cluster + NAD+ + H+ + Na+[side 1]
-
 Vibrio cholerae serotype O1 2 oxidized ferredoxin [iron-sulfur] cluster + NADH + Na+[side 2]
-
 ?
2 reduced ferredoxin [iron-sulfur] cluster + NAD+ + H+ + Na+[side 1] Rnf's electron donation from NADH corresponds to inward flow of Na+ Vibrio cholerae serotype O1 2 oxidized ferredoxin [iron-sulfur] cluster + NADH + Na+[side 2]
-
 ?
2 reduced ferredoxin [iron-sulfur] cluster + NAD+ + H+ + Na+[side 1]
-
 Vibrio cholerae serotype O1 O395N-1 2 oxidized ferredoxin [iron-sulfur] cluster + NADH + Na+[side 2]
-
 ?
2 reduced ferredoxin [iron-sulfur] cluster + NAD+ + H+ + Na+[side 1] Rnf's electron donation from NADH corresponds to inward flow of Na+ Vibrio cholerae serotype O1 O395N-1 2 oxidized ferredoxin [iron-sulfur] cluster + NADH + Na+[side 2]
-
 ?
additional information the recombinant isolated RnfG produces a neutral semiquinone intermediate. The semiquinone species disappeared upon full reduction and is not observed in the denatured protein Vibrio cholerae serotype O1 ?
-
 ?
additional information the recombinant isolated RnfG produces a neutral semiquinone intermediate. The semiquinone species disappeared upon full reduction and is not observed in the denatured protein Vibrio cholerae serotype O1 O395N-1 ?
-
 ?

Subunits

Subunits Comment Organism
hexamer six subunits, RnfA, RnfB, RnfC, RnfD, RnfG, and RnfE, that include several conserved sequence motifs indicating the binding of redox cofactors: RnfC has an NADH binding motif, RnfB and RnfC each contain the sequence motif for an iron-sulfur center Vibrio cholerae serotype O1
monomer 1 * 26000, recombinant isolated subunit RnfG, SDS-PAGE, 1 * 39000, recombinant isolated subunit RnfD, SDS-PAGE Vibrio cholerae serotype O1

Synonyms

Synonyms Comment Organism
Rnf complex
-
 Vibrio cholerae serotype O1

Cofactor

Cofactor Comment Organism Structure
Ferredoxin
-
 Vibrio cholerae serotype O1
flavin flavin covalently bound Thr187 in subunit RnfD. In RnfD, the flavin is bound, not to the SGAT sequence but to the final residues of a TMAT sequence, another variant of the flavin binding motif Vibrio cholerae serotype O1
FMN covalently bound to Thr175 in subunit RnfG, the final threonine of the S(T)GAT sequence Vibrio cholerae serotype O1
additional information subunits RnfD and RnfG both contain the same motif, suggesting that they bind flavins in a similar way, mass spectroscopic analysis, overview. The flavins in RnfG and RnfD are localized in the periplasmic space Vibrio cholerae serotype O1
NAD+ Rnf's electron donation from NADH corresponds to inward flow of Na+. Subunit RnfC has an NADH binding motif Vibrio cholerae serotype O1

General Information

General Information Comment Organism
physiological function enzymes of the Rnf family are bacterial redox-driven ion pumps, coupling an oxidoreduction process to the translocation of Na+ across the cell membrane Vibrio cholerae serotype O1