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.
biliverdin Ixalpha + reduced ferredoxin
PcyA catalyzes the multi-step reduction of biliverdin IXalpha to produce 3Z/3E-phycocyanobilin
724131
Synechocystis sp.
(3Z)-phycocyanobilin + oxidized ferredoxin
?
biliverdin Ixalpha + reduced ferredoxin
recombinant Synechocystis sp. PCC7002 ferredoxin
724131
Synechocystis sp.
(3Z)-phycocyanobilin + oxidized ferredoxin
?
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.
biliverdin Ixalpha + reduced ferredoxin
PcyA catalyzes the multi-step reduction of biliverdin IXalpha to produce 3Z/3E-phycocyanobilin
724131
Synechocystis sp.
(3Z)-phycocyanobilin + oxidized ferredoxin
?
biliverdin Ixalpha + reduced ferredoxin
recombinant Synechocystis sp. PCC7002 ferredoxin
724131
Synechocystis sp.
(3Z)-phycocyanobilin + oxidized ferredoxin
?
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.
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.
742318
Ceron-Carrasco
-
First computational step towa ...
Synechocystis sp. PCC 6803
Comput. Theoret. Chem.
1077
58-64
2016
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742524
Hagiwara
Atomic-resolution structure o ...
Synechocystis sp. PCC 6803
FEBS Lett.
590
3425-3434
2016
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724131
Kabasakal
His74 conservation in the bili ...
Synechocystis sp.
Arch. Biochem. Biophys.
537
233-242
2013
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724317
Alvey
Attachment of noncognate chrom ...
Synechococcus sp., Synechococcus sp. 7002
Biochemistry
50
4890-4902
2011
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725252
Alvey
Effects of modified phycobilin ...
Synechococcus sp.
J. Bacteriol.
193
1663-1671
2011
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711117
Wada
One residue substitution in Pc ...
Synechocystis sp.
Biochem. Biophys. Res. Commun.
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373-377
2010
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712411
Hagiwara
Structural insights into vinyl ...
Synechocystis sp.
J. Biol. Chem.
285
1000-1007
2010
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724287
Shang
Biliverdin amides reveal roles ...
Anabaena sp., Anabaena sp. PCC 7120
Biochemistry
49
6070-6082
2010
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724288
Kohler
Structural basis for hydration ...
Synechocystis sp.
Biochemistry
49
6206-6218
2010
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696870
Zhang
Biosynthesis of fluorescent al ...
Synechocystis sp.
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52
135-140
2009
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698489
Stoll
Structure of the biliverdin ra ...
Nostoc sp. PCC 7120 = FACHB-418, Synechocystis sp.
J. Am. Chem. Soc.
131
1986-1995
2009
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712008
Okada
HO1 and PcyA proteins involved ...
Thermosynechococcus elongatus
FEBS Lett.
583
1251-1256
2009
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685125
Tu
Insight into the radical mecha ...
Nostoc sp. PCC 7120 = FACHB-418
Biochemistry
46
1484-1494
2007
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673636
Mukougawa
Metabolic engineering to produ ...
Synechococcus sp., Synechococcus sp. W8020
FEBS Lett.
580
1333-1338
2006
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1
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673657
Hagiwara
Induced-fitting and electrosta ...
Synechocystis sp.
FEBS Lett.
580
3823-3828
2006
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674669
Tu
A conserved histidine-aspartat ...
Nostoc sp., Nostoc sp. PCC 7120
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281
3127-3136
2006
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1
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674771
Zhao
Chromophore attachment to phyc ...
Anabaena sp., Anabaena sp. PCC 7120
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Hagiwara
Crystal structure of phycocyan ...
Synechocystis sp.
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Tu
Biliverdin reduction by cyanob ...
Anabaena sp.
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657164
Kami
Complementation of phytochrome ...
Synechocystis sp.
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1099-1104
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656197
Frankenberg
Phycocyanobilin:ferredoxin oxi ...
Anabaena sp.
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437763
Tooley
Biosynthesis of the cyanobacte ...
Anabaena sp.
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437673
Frankenberg
Functional genomic analysis of ...
Anabaena sp., Nostoc punctiforme, Prochlorococcus sp., Prochlorococcus sp. CCMP1378 / MED4, Synechococcus sp., Synechococcus sp. WH8102, Synechocystis sp.
Plant Cell
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437764
Tooley
Biosynthesis of a fluorescent ...
Synechocystis sp.
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10560-10565
2001
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437765
Wu
Phycocyanobilin is the natural ...
Mesotaenium caldariorum
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1997
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