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Literature summary for 1.18.1.1 extracted from

  • Hagelueken, G.; Wiehlmann, L.; Adams, T.M.; Kolmar, H.; Heinz, D.W.; Tuemmler, B.; Schubert, W.D.
    Crystal structure of the electron transfer complex rubredoxin-rubredoxin reductase of Pseudomonas aeruginosa (2007), Proc. Natl. Acad. Sci. USA, 104, 12276-12281.
    View publication on PubMedView publication on EuropePMC

Cloned(Commentary)

Cloned (Comment) Organism
gene rdxR or PA5349, expression of His-tagged wild-type enzyme in Escherichia coli Pseudomonas aeruginosa

Crystallization (Commentary)

Crystallization (Comment) Organism
purified recombinant His-tagged enzyme, free or in complex with substrate rubredoxin, sitting drop vapour diffusion method, 20°C, 8.5 mg/ml protein in 100 mM NaCl, 50 mM Tris-HCl, pH 8.0, FAD, and 5 mM 2-mercaptoethanol, in presence or absence of rubredoxin in a 1.2 molar excess, mixing with an equal volume of reservoir solution containing 5% PEG 1000, 40% PEG 300, 0.1 M Tris-HCl, pH 7.0, mother liquor supplemented with 25% PEG 400 is used for cryoprotection, X-ray diffraction structure determination and analysis at 2.3-2.4 A resolution Pseudomonas aeruginosa

Protein Variants

Protein Variants Comment Organism
additional information a mutant of Pseudomonas aeruginosa with an isogenic rdxR transposon insertion is unable to grow on hexadecane Pseudomonas aeruginosa

Metals/Ions

Metals/Ions Comment Organism Structure
additional information the electron transfer between the enzyme and rubredoxin involves a redox active Fe3+, which can be substituted for Ni2+, mechanism, overview Pseudomonas aeruginosa

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
reduced rubredoxin + NAD(P)+ Pseudomonas aeruginosa the enzyme and two rubredoxins form a system indipensable for metabolizing n-alkanes, they constitute an electron transport pathway that shuttles reducing equivalents from carbon metabolism to the membrane-bound alkane hydroxylases AlkB1 and AlkB2 oxidized rubredoxin + NAD(P)H
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Organism

Organism UniProt Comment Textmining
Pseudomonas aeruginosa Q9HTK9 strains TBCF10839 and PAO1, gene PA5349 or rdxR
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Purification (Commentary)

Purification (Comment) Organism
recombinant His-tagged wild-type enzyme from Escherichia coli by nickel affinity and anion exchange chromatography, and gel filtration Pseudomonas aeruginosa

Reaction

Reaction Comment Organism Reaction ID
reduced rubredoxin + NAD+ + H+ = oxidized rubredoxin + NADH the electron transfer from NAD(P)H to rubredoxin involves a reductive and an oxidative step with respect to the enzyme Pseudomonas aeruginosa

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
reduced rubredoxin + NAD(P)+ the enzyme and two rubredoxins form a system indipensable for metabolizing n-alkanes, they constitute an electron transport pathway that shuttles reducing equivalents from carbon metabolism to the membrane-bound alkane hydroxylases AlkB1 and AlkB2 Pseudomonas aeruginosa oxidized rubredoxin + NAD(P)H
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reduced rubredoxin + NAD(P)+ rubredoxin-rubredoxin reductase complex formation, only a small number of direct interactions govern mutual recognition of RdxR and Rdx, corroborating the transient nature of the complex, overview, substrate binding structure and mechanism, the enzyme discriminates between two types of rubredoxins, RubA2 and RubA1, overview Pseudomonas aeruginosa oxidized rubredoxin + NAD(P)H
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Subunits

Subunits Comment Organism
dimer RdxR consists of two cofactor-binding domains and a C-terminal domain essential for the specific recognition of Rdx, crystal structure analysis, dimerization restricts access to the NAD(P)H binding pocket and results in a steric clash between the modeled adenine moiety of NAD(P)H and alpha-helix alpha8' of the neighboring molecule, RdxR dimers form at high protein concentrations used during crystallization, rather than being functionally relevant, overview Pseudomonas aeruginosa

Synonyms

Synonyms Comment Organism
NAD(P)H:rubredoxin reductase
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Pseudomonas aeruginosa
RdxR
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Pseudomonas aeruginosa
rubredoxin reductase
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Pseudomonas aeruginosa

Cofactor

Cofactor Comment Organism Structure
FAD dependent on, binding site structure, overview Pseudomonas aeruginosa
NAD(P)+ binding site structure, overview Pseudomonas aeruginosa