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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.
    View publication on PubMed

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
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?

Organism

EC Number Organism UniProt Comment Textmining
1.3.7.5 Synechocystis sp.
-
gene pycA
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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
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?
1.3.7.5 biliverdin Ixalpha + reduced ferredoxin recombinant Synechocystis sp. PCC7002 ferredoxin Synechocystis sp. (3Z)-phycocyanobilin + oxidized ferredoxin
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?

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
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Synechocystis sp.
1.3.7.5 phycocyanobilin:ferredoxin oxidoreductase
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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.