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1.13.11.3: protocatechuate 3,4-dioxygenase

This is an abbreviated version!
For detailed information about protocatechuate 3,4-dioxygenase, go to the full flat file.

Word Map on EC 1.13.11.3

Reaction

3,4-dihydroxybenzoate
+
O2
=
3-carboxy-cis,cis-muconate

Synonyms

3,4-PCase, 3,4-PCD, 3,4-PCDase, 3,4-POD, EC 1.13.1.3, EC 1.99.2.3, More, oxygenase, protocatechuate 3,4-di-, P3,4DO, P3,4O enzyme, P34O, PCA 3,4-dioxygenase, PcaG, PcaH, PcaHG, PCD, protocatchetuate 3,4-dioxygenase, protocatechuate 3,4-dioxygenase, protocatechuate oxygenase, protocatechuic 3,4-dioxygenase, protocatechuic 3,4-oxygenase, protocatechuic acid oxidase

ECTree

     1 Oxidoreductases
         1.13 Acting on single donors with incorporation of molecular oxygen (oxygenases)
             1.13.11 With incorporation of two atoms of oxygen
                1.13.11.3 protocatechuate 3,4-dioxygenase

Crystallization

Crystallization on EC 1.13.11.3 - protocatechuate 3,4-dioxygenase

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CRYSTALLIZATION (Commentary)
ORGANISM
UNIPROT
LITERATURE
recombinant enzyme, hanging drop method with 1.8 M ammonium sulfate, 50 mM Tris-HCl pH 7.0 in the reservoir and 10 mg/ml enzyme in the drop at 18°C, crystals grow as perfect dodecahedrons, crystal structure with 2.2 A resolution
-
microdialysis of approx. 0.05 ml of enzyme solution, 80 mg/ml against 10 ml volumes of buffered solutions, large single crystals
-
vapor equilibration at room temperature from solutions containing potassium phosphate pH 7.5, 1.08-1.2 M reservoir concentrations, initial enzyme concentration 6.7-10.5 mg/ml
-
addition of 0.1 ml 1 M 2-mercaptoethanol to 10 ml enzyme solution in 50 mM Tris buffer pH 8.5, rhombic crystals appear after 1 week in a refrigerator, crystallization is also achieved by adding 1 mM iodoacetamide instead of mercaptoethanol or by dialyzing the enzyme against cold, destilled water for 2 days
-
X-ray crystallography
-
free interface diffusion cell, equal volumes of concentrated enzyme and ammonium sulfate solutions ranging from 37-47% saturation, 4°C, 2.5 A resolution
-
vapor diffusion in hanging drops, crystal structure at 2.1-2.2 A resolution
-
vapor diffusion with ammonium sulfate as the precipitant at 4°C, recombinant wild-type and Y447H mutant enzyme
-