Any feedback?
Please rate this page
(literature.php)
(0/150)

BRENDA support

Literature summary for 1.3.7.7 extracted from

  • Broecker, M.J.; Schomburg, S.; Heinz, D.W.; Jahn, D.; Schubert, W.D.; Moser, J.
    Crystal structure of the nitrogenase-like dark operative protochlorophyllide oxidoreductase catalytic complex (ChlN/ChlB)2 (2010), J. Biol. Chem., 285, 27336-27345.
    View publication on PubMedView publication on EuropePMC

Cloned(Commentary)

Cloned (Comment) Organism
expression of N-terminally GST-tagged (ChlN/ChlB)2 complex Thermosynechococcus vestitus

Crystallization (Commentary)

Crystallization (Comment) Organism
native and SeMet-labeled catalytic (ChlN/ChlB)2 complex, hanging drop vapor diffusion, 17°C, mixing of 0.003 ml of 10 mg/ml protein in 100 mM HEPES-NaOH, pH 7.5, 150 mM NaCl, and 10 mM MgCl2, with 0.003 ml of reservoir solution consisting of 9.5% PEG 6000, 85 mM HEPES-NaOH, pH 7.1, 14.3% 2-methyl pentane-2,4-diol, and 15% glycerol as cryoprotectants or 10.5% PEG 6000, 85 mM HEPES-NaOH, pH7.5, 14.3% 2-methyl pentane-2,4-diol, and 15% glycerol as cryoprotectants for selenomethionine-labeled protein, 3-5 days,X-ray diffraction structure determination and analysis at 2.4-2.81 A resolution Thermosynechococcus vestitus

Protein Variants

Protein Variants Comment Organism
C95A inactive protein, probably due to destabilization of the [4Fe-4S] cluster environment Thermosynechococcus vestitus
C95S inactive protein, probably due to destabilization of the [4Fe-4S] cluster environment Thermosynechococcus vestitus

Metals/Ions

Metals/Ions Comment Organism Structure
Fe2+ the enzyme contains intersubunit [4Fe-4S] cluster in the homodimer formed by two ChlL subunits. The [4Fe-4S] cluster coordinated by an aspartate oxygen alongside three cysteine ligands, structure, overview Thermosynechococcus vestitus
Mg2+
-
Thermosynechococcus vestitus

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
chlorophyllide a + reduced ferredoxin + 2 ATP + 2 H2O Thermosynechococcus vestitus
-
protochlorophyllide + oxidized ferredoxin + 2 ADP + 2 phosphate + 2 H+
-
?

Organism

Organism UniProt Comment Textmining
Thermosynechococcus vestitus
-
-
-

Purification (Commentary)

Purification (Comment) Organism
recombinant N-terminally GST-tagged (ChlN/ChlB)2 complex by glutathione affinity chromatography, and gel filtration and ultrafiltration Thermosynechococcus vestitus

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
chlorophyllide a + reduced ferredoxin + 2 ATP + 2 H2O
-
Thermosynechococcus vestitus protochlorophyllide + oxidized ferredoxin + 2 ADP + 2 phosphate + 2 H+
-
?
chlorophyllide a + reduced ferredoxin + 2 ATP + 2 H2O ferredoxin provides a single electron to ChlL2, which in turn transfers an electron to (ChlN/ChlB)2. Hydrolysis of the two ATP molecules results in the dissociation of ChlL2 from reduced (ChlN/ChlB)2. Protochlorophyllide reduction is completed after two sequential catalytic redox cycles. Substrate recognition by (ChlN/ChlB)2 essentially involves all functional groups of the substrate, modeling of the substrate binding site of (ChlN/ChlB)2, overview. Electron transfer pathway via the various redox centers of DPOR to the substrate, overview Thermosynechococcus vestitus protochlorophyllide + oxidized ferredoxin + 2 ADP + 2 phosphate + 2 H+
-
?
additional information the invitro assay is performed with purified recombinant GST-tagged (ChlN/ChlB)2 complex and a ChlL2 subunit purified from Prochlorococcus marinus Thermosynechococcus vestitus ?
-
?

Subunits

Subunits Comment Organism
heterooctamer (alpha2)2(betagamma)4, DPOR is composed of the subunits ChlL, ChlN, and ChlB. Homodimeric ChlL2 bearing an intersubunit [4Fe-4S] cluster is an ATP-dependent reductase transferring single electrons to the heterotetrameric (ChlN/ChlB)2 complex. The latter contains two intersubunit [4Fe-4S] clusters and two protochlorophyllide binding sites, respectively, structure analysis of the catalytic (ChlN/ChlB)2 complex, overview. Subunits ChlN and ChlB exhibit a related architecture of three subdomains each built around a central, parallel beta-sheet surrounded by alpha-helices. Two ChlL2 dimers simultaneously interact with the (ChlN/ChlB)2 tetramer, giving rise to a heterooctameric holoenzyme Thermosynechococcus vestitus
More model of the complete hetero-octameric DPOR and distance between cofactors, overview Thermosynechococcus vestitus

Synonyms

Synonyms Comment Organism
dark operative protochlorophyllide oxidoreductase
-
Thermosynechococcus vestitus
DPOR
-
Thermosynechococcus vestitus

Cofactor

Cofactor Comment Organism Structure
ATP dependent on, the homodimeric ChlL2 bearing an intersubunit [4Fe-4S] cluster is an ATP-dependent reductase Thermosynechococcus vestitus
Ferredoxin the enzyme contains intersubunit [4Fe-4S] cluster in the homodimer formed by two ChlL subunits. The [4Fe-4S] cluster coordinated by an aspartate oxygen alongside three cysteine ligands, structure, overview. Iron-sulfur cluster coordination in DPOR compared to nitrogenase, overview Thermosynechococcus vestitus