BRENDA - Reference to 1.3.7.5; Id = 724131

gene pycA, expression of wild-type and mutant enzymes in Escherichia coli strain BL21(DE3)

Synechocystis sp.

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.

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.

biliverdin Ixalpha + reduced ferredoxin

Synechocystis sp.

PcyA catalyzes the multi-step reduction of biliverdin IXalpha to produce 3Z/3E-phycocyanobilin

(3Z)-phycocyanobilin + oxidized ferredoxin

-

?

Synechocystis sp.

-

gene pycA

-

recombinant wild-type and mutant enzymes from Escherichia coli strain Bl21(DE3)

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

-

?

More

structure analysis of recombinant wild-type and mutant enzymes, overview

Synechocystis sp.

PcyA

-

Synechocystis sp.

phycocyanobilin:ferredoxin oxidoreductase

-

Synechocystis sp.

16

-

assay at

Synechocystis sp.

8.5

-

assay at

Synechocystis sp.

Ferredoxin

-

Synechocystis sp.

gene pycA, expression of wild-type and mutant enzymes in Escherichia coli strain BL21(DE3)

Synechocystis sp.

Ferredoxin

-

Synechocystis sp.

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.

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.

biliverdin Ixalpha + reduced ferredoxin

Synechocystis sp.

PcyA catalyzes the multi-step reduction of biliverdin IXalpha to produce 3Z/3E-phycocyanobilin

(3Z)-phycocyanobilin + oxidized ferredoxin

-

?

recombinant wild-type and mutant enzymes from Escherichia coli strain Bl21(DE3)

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

-

?

More

structure analysis of recombinant wild-type and mutant enzymes, overview

Synechocystis sp.

16

-

assay at

Synechocystis sp.

8.5

-

assay at

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.

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

-

1

-

1

-

1

-

742524

Hagiwara

Atomic-resolution structure o ...

Synechocystis sp. PCC 6803

FEBS Lett.

590

3425-3434

2016

-

1

-

2

-

1

-

1

-

1

-

724131

Kabasakal

His74 conservation in the bili ...

Synechocystis sp.

Arch. Biochem. Biophys.

537

233-242

2013

-

1

3

-

1

-

1

-

1

-

2

1

2

1

-

1

-

1

-

1

3

-

1

-

1

-

2

1

-

1

-

3

-

724317

Alvey

Attachment of noncognate chrom ...

Synechococcus sp., Synechococcus sp. 7002

Biochemistry

50

4890-4902

2011

-

1

-

2

-

10

-

2

-

2

-

1

-

1

-

2

-

2

-

1

-

725252

Alvey

Effects of modified phycobilin ...

Synechococcus sp.

J. Bacteriol.

193

1663-1671

2011

-

1

-

2

-

4

-

4

-

3

-

1

-

1

-

1

-

2

-

4

-

3

-

711117

Wada

One residue substitution in Pc ...

Synechocystis sp.

Biochem. Biophys. Res. Commun.

402

373-377

2010

-

1

-

1

-

3

-

2

-

1

-

1

-

1

-

1

-

2

-

712411

Hagiwara

Structural insights into vinyl ...

Synechocystis sp.

J. Biol. Chem.

285

1000-1007

2010

-

1

-

1

-

5

-

3

-

1

-

1

-

1

-

1

-

1

-

3

-

1

-

1

-

724287

Shang

Biliverdin amides reveal roles ...

Anabaena sp., Anabaena sp. PCC 7120

Biochemistry

49

6070-6082

2010

-

1

-

2

-

20

-

1

-

13

-

2

-

2

-

1

-

1

-

2

-

1

-

13

-

2

-

2

-

724288

Kohler

Structural basis for hydration ...

Synechocystis sp.

Biochemistry

49

6206-6218

2010

-

1

2

-

1

-

3

-

1

-

1

-

1

-

1

2

-

1

-

1

-

1

-

2

-

696870

Zhang

Biosynthesis of fluorescent al ...

Synechocystis sp.

Biotechnol. Appl. Biochem.

52

135-140

2009

-

1

-

1

-

1

-

1

-

1

-

698489

Stoll

Structure of the biliverdin ra ...

Nostoc sp. PCC 7120 = FACHB-418, Synechocystis sp.

J. Am. Chem. Soc.

131

1986-1995

2009

-

2

3

-

7

-

2

3

-

712008

Okada

HO1 and PcyA proteins involved ...

Thermosynechococcus elongatus

FEBS Lett.

583

1251-1256

2009

-

1

-

1

-

1

-

1

-

1

2

1

-

1

-

1

-

1

-

1

-

1

-

1

-

1

-

685125

Tu

Insight into the radical mecha ...

Nostoc sp. PCC 7120 = FACHB-418

Biochemistry

46

1484-1494

2007

-

1

5

-

4

-

1

-

1

-

2

-

1

5

-

1

-

1

-

673636

Mukougawa

Metabolic engineering to produ ...

Synechococcus sp., Synechococcus sp. W8020

FEBS Lett.

580

1333-1338

2006

-

1

-

2

-

2

-

1

-

1

-

2

-

673657

Hagiwara

Induced-fitting and electrosta ...

Synechocystis sp.

FEBS Lett.

580

3823-3828

2006

-

1

-

4

-

1

-

1

-

674669

Tu

A conserved histidine-aspartat ...

Nostoc sp., Nostoc sp. PCC 7120

J. Biol. Chem.

281

3127-3136

2006

-

1

-

16

-

1

-

15

-

1

-

1

2

-

1

-

1

-

16

-

1

-

1

-

1

2

-

674771

Zhao

Chromophore attachment to phyc ...

Anabaena sp., Anabaena sp. PCC 7120

J. Biol. Chem.

281

8573-8581

2006

-

1

-

18

-

2

-

1

-

1

-

2

-

676847

Hagiwara

Crystal structure of phycocyan ...

Synechocystis sp.

Proc. Natl. Acad. Sci. USA

103

27-32

2006

-

1

-

3

-

1

-

1

-

1

-

1

-

1

-

655796

Tu

Biliverdin reduction by cyanob ...

Anabaena sp.

J. Am. Chem. Soc.

126

8682-8693

2004

-

1

-

1

-

2

-

1

-

5

-

1

-

1

-

1

-

1

-

5

-

657164

Kami

Complementation of phytochrome ...

Synechocystis sp.

Proc. Natl. Acad. Sci. USA

101

1099-1104

2004

-

1

-

1

-

3

-

1

-

1

-

1

-

1

-

1

-

656197

Frankenberg

Phycocyanobilin:ferredoxin oxi ...

Anabaena sp.

J. Biol. Chem.

278

9219-9226

2003

-

1

-

2

1

-

5

-

4

1

-

1

-

2

1

-

4

1

-

437763

Tooley

Biosynthesis of the cyanobacte ...

Anabaena sp.

J. Bacteriol.

184

4666-4671

2002

-

1

-

1

-

1

-

1

-

1

-

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

13

965-978

2001

-

5

-

2

-

8

-

5

-

7

-

5

-

2

-

5

-

7

-

437764

Tooley

Biosynthesis of a fluorescent ...

Synechocystis sp.

Proc. Natl. Acad. Sci. USA

98

10560-10565

2001

-

1

-

4

-

1

-

1

-

1

-

437765

Wu

Phycocyanobilin is the natural ...

Mesotaenium caldariorum

J. Biol. Chem.

272

25700-25705

1997

-

1

-

1

-

1

-

1

-

1

-

His74 conservation in the bilin reductase PcyA family reflects an important role in protein-substrate structure and dynamics

Data extracted from this reference: