Cloned (Comment) | Organism |
---|---|
gene pycA, expression of wild-type and mutant enzymes in Escherichia coli strain BL21(DE3) | Synechocystis sp. |
Crystallization (Comment) | Organism |
---|---|
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 | Comment | Organism |
---|---|---|
H74A | site-directed mutagenesis, inactive mutant | Synechocystis sp. |
H74E | site-directed mutagenesis, the mutant retains reasonable activity | Synechocystis sp. |
H74Q | site-directed mutagenesis, the mutant retains reasonable activity | Synechocystis sp. |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
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 | UniProt | Comment | Textmining |
---|---|---|---|
Synechocystis sp. | - |
gene pycA | - |
Purification (Comment) | Organism |
---|---|
recombinant wild-type and mutant enzymes from Escherichia coli strain Bl21(DE3) | Synechocystis sp. |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
biliverdin Ixalpha + reduced ferredoxin | PcyA catalyzes the multi-step reduction of biliverdin IXalpha to produce 3Z/3E-phycocyanobilin | Synechocystis sp. | (3Z)-phycocyanobilin + oxidized ferredoxin | - |
? | |
biliverdin Ixalpha + reduced ferredoxin | recombinant Synechocystis sp. PCC7002 ferredoxin | Synechocystis sp. | (3Z)-phycocyanobilin + oxidized ferredoxin | - |
? |
Subunits | Comment | Organism |
---|---|---|
More | structure analysis of recombinant wild-type and mutant enzymes, overview | Synechocystis sp. |
Synonyms | Comment | Organism |
---|---|---|
PcyA | - |
Synechocystis sp. |
phycocyanobilin:ferredoxin oxidoreductase | - |
Synechocystis sp. |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
16 | - |
assay at | Synechocystis sp. |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
8.5 | - |
assay at | Synechocystis sp. |
Cofactor | Comment | Organism | Structure |
---|---|---|---|
Ferredoxin | - |
Synechocystis sp. |
General Information | Comment | Organism |
---|---|---|
evolution | the enzyme is a member of the ferredoxin-dependent biliverdin reductase (FDBR) family | Synechocystis sp. |
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. |
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. |