Literature summary extracted from
Kabasakal, B.; Gae, D.D.; Li, J.; Clark Lagarias, J.; Koehl, P.; Fisher, A.J.
His74 conservation in the bilin reductase PcyA family reflects an important role in protein-substrate structure and dynamics (2013), Arch. Biochem. Biophys., 537, 233-242.
Cloned(Commentary)
EC Number |
Cloned (Comment) |
Organism |
---|
1.3.7.5 |
gene pycA, expression of wild-type and mutant enzymes in Escherichia coli strain BL21(DE3) |
Synechocystis sp. |
Crystallization (Commentary)
EC Number |
Crystallization (Comment) |
Organism |
---|
1.3.7.5 |
purified recombinant wild-type and mutant enzymes, hanging drop vapor diffusion method, mixing of 0.002 ml of protein solution containing 15 mg/mL protein and 0.67 mM biliverdin IXalpha, with 0.002 ml of reservoir solution containing 1-1.25 M sodium citrate, 0.1-0.4 M NaCl, and 0.1 M Tris HCl, pH 7.0, 21°C, 1-2 weeks. Crystal trials are set up under green safelight and stored in the dark, X-ray diffraction structure determination and analysis at 1.18-1.49 A resolution |
Synechocystis sp. |
Protein Variants
EC Number |
Protein Variants |
Comment |
Organism |
---|
1.3.7.5 |
H74A |
site-directed mutagenesis, inactive mutant |
Synechocystis sp. |
1.3.7.5 |
H74E |
site-directed mutagenesis, the mutant retains reasonable activity |
Synechocystis sp. |
1.3.7.5 |
H74Q |
site-directed mutagenesis, the mutant retains reasonable activity |
Synechocystis sp. |
Natural Substrates/ Products (Substrates)
EC Number |
Natural Substrates |
Organism |
Comment (Nat. Sub.) |
Natural Products |
Comment (Nat. Pro.) |
Rev. |
Reac. |
---|
1.3.7.5 |
biliverdin Ixalpha + reduced ferredoxin |
Synechocystis sp. |
PcyA catalyzes the multi-step reduction of biliverdin IXalpha to produce 3Z/3E-phycocyanobilin |
(3Z)-phycocyanobilin + oxidized ferredoxin |
- |
? |
|
Organism
EC Number |
Organism |
UniProt |
Comment |
Textmining |
---|
1.3.7.5 |
Synechocystis sp. |
- |
gene pycA |
- |
Purification (Commentary)
EC Number |
Purification (Comment) |
Organism |
---|
1.3.7.5 |
recombinant wild-type and mutant enzymes from Escherichia coli strain Bl21(DE3) |
Synechocystis sp. |
Substrates and Products (Substrate)
EC Number |
Substrates |
Comment Substrates |
Organism |
Products |
Comment (Products) |
Rev. |
Reac. |
---|
1.3.7.5 |
biliverdin Ixalpha + reduced ferredoxin |
PcyA catalyzes the multi-step reduction of biliverdin IXalpha to produce 3Z/3E-phycocyanobilin |
Synechocystis sp. |
(3Z)-phycocyanobilin + oxidized ferredoxin |
- |
? |
|
1.3.7.5 |
biliverdin Ixalpha + reduced ferredoxin |
recombinant Synechocystis sp. PCC7002 ferredoxin |
Synechocystis sp. |
(3Z)-phycocyanobilin + oxidized ferredoxin |
- |
? |
|
Subunits
EC Number |
Subunits |
Comment |
Organism |
---|
1.3.7.5 |
More |
structure analysis of recombinant wild-type and mutant enzymes, overview |
Synechocystis sp. |
Synonyms
EC Number |
Synonyms |
Comment |
Organism |
---|
1.3.7.5 |
PcyA |
- |
Synechocystis sp. |
1.3.7.5 |
phycocyanobilin:ferredoxin oxidoreductase |
- |
Synechocystis sp. |
Temperature Optimum [°C]
EC Number |
Temperature Optimum [°C] |
Temperature Optimum Maximum [°C] |
Comment |
Organism |
---|
1.3.7.5 |
16 |
- |
assay at |
Synechocystis sp. |
pH Optimum
EC Number |
pH Optimum Minimum |
pH Optimum Maximum |
Comment |
Organism |
---|
1.3.7.5 |
8.5 |
- |
assay at |
Synechocystis sp. |
Cofactor
EC Number |
Cofactor |
Comment |
Organism |
Structure |
---|
1.3.7.5 |
Ferredoxin |
- |
Synechocystis sp. |
|
General Information
EC Number |
General Information |
Comment |
Organism |
---|
1.3.7.5 |
evolution |
the enzyme is a member of the ferredoxin-dependent biliverdin reductase (FDBR) family |
Synechocystis sp. |
1.3.7.5 |
additional information |
the fully conserved residue His74 plays a critical role in the H-bonding network that permits proton transfer, molecular dynamics simulations, overview. A conserved buried water molecule that bridges His74 and catalytically essential His88 is not required for activity. The crucial active site residue Asp105 is more dynamic in H74A compared to wild-type PcyA and the two other His74 variants, supporting the conclusion that the Ala74 mutation has increased the flexibility of the active site. Structure analysis of recombinant wild-type and mutant enzymes, overview |
Synechocystis sp. |
1.3.7.5 |
physiological function |
bilin reductase PcyA catalyzes the proton-coupled four-electron reduction of biliverdin IXa's two vinyl groups to produce phycocyanobilin, an essential chromophore for phytochromes, cyanobacteriochromes and phycobiliproteins |
Synechocystis sp. |