EC Number | Cloned (Comment) | Organism |
---|---|---|
1.3.7.7 | expressed in Escherichia coli JM105 cells | Rhodobacter capsulatus |
1.3.7.7 | protochlorophyllide-bound and protochlorophyllide-free forms of NB-protein are overexpressed in Rhodobacter capsulatus strain DB176 and Escherichia coli strain JM105 | Rhodobacter capsulatus |
EC Number | Crystallization (Comment) | Organism |
---|---|---|
1.3.7.7 | hanging drop vapor diffusion method, the protochlorophyllide-bound form of NB-protein is crystallized using 200 mM sodium/potassium phosphate buffer (pH 5.0) containing 5 mM dithiothreitol and 10% (w/v) ethylene glycol at 4°C, to which 16% (w/v) and 14% (w/v) PEG4K are added in aerobic and anaerobic conditions, respectively, as precipitants. Protochlorophyllide -free and selenomethionine-substituted recombinant NB-proteins are crystallized at 20°C using 20% (w/v) PEG3350 containing 200 mM ammonium chloride and 5mM dithiothreitol. D36C and D36A variants are crystallized at 4°C using 20% (w/v) PEG3350 containing 200 mM sodium chloride, 100 mM MOPS/NaOH (pH 7.0) and 5 mM dithiothreitol as a precipitant | Rhodobacter capsulatus |
1.3.7.7 | purified recombinant protochlorophyllide-bound and protochlorophyllide-free forms of NB-protein of DPOR, and purified recombinant selenomethionine-substituted protochlorophyllide-free forms of mutants D36A and D36C, hanging-drop vapour diffusion method., X-ray diffraction structure determination and analysis at 2.3-2.9 A resolution | Rhodobacter capsulatus |
EC Number | Protein Variants | Comment | Organism |
---|---|---|---|
1.3.7.7 | C112A | inactive | Rhodobacter capsulatus |
1.3.7.7 | C26A | inactive | Rhodobacter capsulatus |
1.3.7.7 | C51A | the mutation almost abolishes the activity of the enzyme (less than 5% compared to the wild type enzyme) | Rhodobacter capsulatus |
1.3.7.7 | C95A | inactive | Rhodobacter capsulatus |
1.3.7.7 | D36A | site-directed mutagenesis, mutation in BchB, mutant NB-cluster structure compered to the wild-type enzyme | Rhodobacter capsulatus |
1.3.7.7 | D36A | the mutant exhibits low activity (13% compared to the wild type enzyme) | Rhodobacter capsulatus |
1.3.7.7 | D36C | site-directed mutagenesis, mutation in BchB, mutant NB-cluster structure compered to the wild-type enzyme | Rhodobacter capsulatus |
1.3.7.7 | D36C | the mutation almost abolishes the activity of the enzyme (less than 5% compared to the wild type enzyme) | Rhodobacter capsulatus |
1.3.7.7 | D36S | site-directed mutagenesis, mutation in BchB, mutant NB-cluster structure compered to the wild-type enzyme | Rhodobacter capsulatus |
1.3.7.7 | D36S | the mutation almost abolishes the activity of the enzyme (less than 5% compared to the wild type enzyme) | Rhodobacter capsulatus |
EC Number | Inhibitors | Comment | Organism | Structure |
---|---|---|---|---|
1.3.7.7 | chlorophyll c | competitive inhibitor | Rhodobacter capsulatus |
EC Number | Metals/Ions | Comment | Organism | Structure |
---|---|---|---|---|
1.3.7.7 | Fe2+ | iron-sulfur cluster in the NB protein | Rhodobacter capsulatus | |
1.3.7.7 | Mg2+ | - |
Rhodobacter capsulatus |
EC Number | Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|---|
1.3.7.7 | chlorophyllide a + reduced ferredoxin + 2 ATP + 2 H2O | Rhodobacter capsulatus | - |
protochlorophyllide + oxidized ferredoxin + 2 ADP + 2 phosphate + 2 H+ | - |
? | |
1.3.7.7 | chlorophyllide a + reduced ferredoxin + 2 ATP + 2 H2O | Rhodobacter capsulatus DB176 | - |
protochlorophyllide + oxidized ferredoxin + 2 ADP + 2 phosphate + 2 H+ | - |
? |
EC Number | Organism | UniProt | Comment | Textmining |
---|---|---|---|---|
1.3.7.7 | Rhodobacter capsulatus | - |
- |
- |
1.3.7.7 | Rhodobacter capsulatus DB176 | - |
- |
- |
EC Number | Purification (Comment) | Organism |
---|---|---|
1.3.7.7 | recombinant protochlorophyllide-bound and protochlorophyllide-free forms of NB-protein from Rhodobacter capsulatus strain DB176 and Escherichia coli strain JM105 by affinity chromatography and gel filtration | Rhodobacter capsulatus |
1.3.7.7 | Strep-tactin column chromatography and Superdex 200 gel filtration | Rhodobacter capsulatus |
EC Number | Reaction | Comment | Organism | Reaction ID |
---|---|---|---|---|
1.3.7.7 | chlorophyllide a + oxidized ferredoxin + 2 ADP + 2 phosphate = protochlorophyllide a + reduced ferredoxin + 2 ATP + 2 H2O | reaction mechanism for trans-specific reduction, overview | Rhodobacter capsulatus |
EC Number | Specific Activity Minimum [µmol/min/mg] | Specific Activity Maximum [µmol/min/mg] | Comment | Organism |
---|---|---|---|---|
1.3.7.7 | 0.026 | - |
purified recombinant protochlorophyllide-bound NB-protein, recovered from crystals, pH not specified in the publication, temperature not specified in the publication | Rhodobacter capsulatus |
1.3.7.7 | 0.05 | - |
purified recombinant protochlorophyllide-free NB-protein, recovered from crystals, pH not specified in the publication, temperature not specified in the publication | Rhodobacter capsulatus |
EC Number | Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|---|
1.3.7.7 | chlorophyllide a + reduced ferredoxin + 2 ATP + 2 H2O | - |
Rhodobacter capsulatus | protochlorophyllide + oxidized ferredoxin + 2 ADP + 2 phosphate + 2 H+ | - |
? | |
1.3.7.7 | chlorophyllide a + reduced ferredoxin + 2 ATP + 2 H2O | - |
Rhodobacter capsulatus DB176 | protochlorophyllide + oxidized ferredoxin + 2 ADP + 2 phosphate + 2 H+ | - |
? | |
1.3.7.7 | chlorophyllide a + reduced ferredoxin + ATP | - |
Rhodobacter capsulatus | protochlorophyllide + oxidized ferredoxin + ADP + phosphate | - |
? | |
1.3.7.7 | chlorophyllide a + reduced ferredoxin + ATP | - |
Rhodobacter capsulatus DB176 | protochlorophyllide + oxidized ferredoxin + ADP + phosphate | - |
? | |
1.3.7.7 | additional information | DPOR is a nitrogenase-like enzyme consisting of two components, L-protein (a BchL dimer) and NB-protein (a BchN-BchB heterotetramer) | Rhodobacter capsulatus | ? | - |
? | |
1.3.7.7 | additional information | each catalytic BchN-BchB unit contains one protochlorophyllidee and one iron-sulfur NB-cluster coordinated uniquely by one aspartate and three cysteines. Unique aspartate ligation is not necessarily needed for the cluster assembly but is essential for the catalytic activity. Specific protochlorophyllide-binding accompanies the partial unwinding of an alpha-helix that belongs to the next catalytic BchN-BchB unit, unique trans-specific reduction mechanism in which the distorted C17-propionate of protochlorophyllid and an aspartate from BchB serve as proton donors for C18 and C17 of protochlorophyllide, respectively, overview | Rhodobacter capsulatus | ? | - |
? | |
1.3.7.7 | additional information | the C175C18 double bond of chlorophyll c is not reduced by DPOR | Rhodobacter capsulatus | ? | - |
? | |
1.3.7.7 | additional information | DPOR is a nitrogenase-like enzyme consisting of two components, L-protein (a BchL dimer) and NB-protein (a BchN-BchB heterotetramer) | Rhodobacter capsulatus DB176 | ? | - |
? | |
1.3.7.7 | additional information | each catalytic BchN-BchB unit contains one protochlorophyllidee and one iron-sulfur NB-cluster coordinated uniquely by one aspartate and three cysteines. Unique aspartate ligation is not necessarily needed for the cluster assembly but is essential for the catalytic activity. Specific protochlorophyllide-binding accompanies the partial unwinding of an alpha-helix that belongs to the next catalytic BchN-BchB unit, unique trans-specific reduction mechanism in which the distorted C17-propionate of protochlorophyllid and an aspartate from BchB serve as proton donors for C18 and C17 of protochlorophyllide, respectively, overview | Rhodobacter capsulatus DB176 | ? | - |
? | |
1.3.7.7 | additional information | the C175C18 double bond of chlorophyll c is not reduced by DPOR | Rhodobacter capsulatus DB176 | ? | - |
? |
EC Number | Subunits | Comment | Organism |
---|---|---|---|
1.3.7.7 | heterooctamer | (alpha2)2(betagamma)4, DPOR is a nitrogenase-like enzyme consisting of two components, L-protein, a BchL dimer, and NB-protein, a BchN-BchB heterotetramer, which are structurally related to nitrogenase Fe protein and MoFe protein, respectively | Rhodobacter capsulatus |
1.3.7.7 | More | each catalytic BchN-BchB unit contains one protochlorophyllide and one iron-sulfur NB-cluster coordinated uniquely by one aspartate and three cysteines. Unique aspartate ligation is not necessarily needed for the cluster assembly but is essential for the catalytic activity. Specific protochlorophyllide-binding accompanies the partial unwinding of an alpha-helix that belongs to the next catalytic BchN-BchB unit, unique trans-specific reduction mechanism in which the distorted C17-propionate of protochlorophyllide and an aspartate from BchB serve as proton donors for C18 and C17 of protochlorophyllide, respectively, overview | Rhodobacter capsulatus |
EC Number | Synonyms | Comment | Organism |
---|---|---|---|
1.3.7.7 | dark-operative Pchlide oxidoreductase | - |
Rhodobacter capsulatus |
1.3.7.7 | DPOR | - |
Rhodobacter capsulatus |
1.3.7.7 | light-independent (dark-operative) Pchlide oxidoreductase | - |
Rhodobacter capsulatus |
1.3.7.7 | light-independent protochlorophyllide reductase | - |
Rhodobacter capsulatus |
EC Number | Cofactor | Comment | Organism | Structure |
---|---|---|---|---|
1.3.7.7 | 4Fe-4S-center | - |
Rhodobacter capsulatus | |
1.3.7.7 | ATP | - |
Rhodobacter capsulatus | |
1.3.7.7 | Ferredoxin | each catalytic BchN-BchB unit contains one protochlorophyllidee and one iron-sulfur NB-cluster coordinated uniquely by one aspartate and three cysteines. Unique aspartate ligation is not necessarily needed for the cluster assembly but is essential for the catalytic activity, overview | Rhodobacter capsulatus |
EC Number | General Information | Comment | Organism |
---|---|---|---|
1.3.7.7 | physiological function | DPOR performs reduction of the C17-C18 double bond of protochlorophyllide to form chlorophyllide a, the direct precursor of chlorophyll a in a light-independent, dark-operative way of action | Rhodobacter capsulatus |