Literature summary for 4.2.1.75 extracted from

Peng, S.; Zhang, H.; Gao, Y.; Pan, X.; Cao, P.; Li, M.; Chang, W.
Crystal structure of uroporphyrinogen III synthase from Pseudomonas syringae pv. tomato DC3000 (2011), Biochem. Biophys. Res. Commun., 408, 576-581.

Cloned(Commentary)

Cloned (Comment)	Organism
gene pspto_0129, expression of His6-tagged enzyme in Escherichia coli strain BL21 (DE3). The selenomethionine-substituted derivative is produced by expression in an Escherichia coli methionine auxotrophic strain B834(DE3)	Pseudomonas syringae

Crystallization (Commentary)

Crystallization (Comment)	Organism
purified wild-type recombinant His6-tagged U3S and selenomethionine-substituted derivative, 0.001 ml of 20 mg/ml protein in 20 mM Hepes, pH 7.2, 20 mM NaCl, is mixed with 0.001 ml of reservoir solution containing 20% PEG 3350, 0.2 M tri-Na citrate, and equilibrated against 0.1 ml of reservoir solution, about a month at 8°C and followed by microseeding method, X-ray diffraction structure determination and analysis at 2.5 A resolution, single wavelength anomalous dispersion method	Pseudomonas syringae

Crystallization (Comment)

Organism

purified wild-type recombinant His6-tagged U3S and selenomethionine-substituted derivative, 0.001 ml of 20 mg/ml protein in 20 mM Hepes, pH 7.2, 20 mM NaCl, is mixed with 0.001 ml of reservoir solution containing 20% PEG 3350, 0.2 M tri-Na citrate, and equilibrated against 0.1 ml of reservoir solution, about a month at 8°C and followed by microseeding method, X-ray diffraction structure determination and analysis at 2.5 A resolution, single wavelength anomalous dispersion method

Pseudomonas syringae

Protein Variants

Protein Variants	Comment	Organism
E114A	site-directed mutagenesis, the mutant shows activity slightly reduced compared to the wild-type enzyme	Pseudomonas syringae
E38A	site-directed mutagenesis, the mutant shows activity similar to the wild-type enzyme	Pseudomonas syringae
L116A	site-directed mutagenesis, the mutant shows activity slightly reduced compared to the wild-type enzyme	Pseudomonas syringae
R164A	site-directed mutagenesis, the mutation causes a decrease in enzymatic activity to about 70% that of the wild-type enzyme	Pseudomonas syringae
R219A	site-directed mutagenesis, the mutation causes a decrease in enzymatic activity to about 25% that of the wild-type enzyme	Pseudomonas syringae

Natural Substrates/ Products (Substrates)

Natural Substrates	Organism	Comment (Nat. Sub.)	Natural Products	Comment (Nat. Pro.)	Rev.	Reac.
hydroxymethylbilane	Pseudomonas syringae	-	uroporphyrinogen III + H2O	-	?
hydroxymethylbilane	Pseudomonas syringae DC3000	-	uroporphyrinogen III + H2O	-	?

Organism

Organism	UniProt	Comment	Textmining
Pseudomonas syringae	Q88B90	pv. tomato, gene pspto_0129	-
Pseudomonas syringae DC3000	Q88B90	pv. tomato, gene pspto_0129	-

Purification (Commentary)

Purification (Comment)	Organism
recombinant His6-tagged enzyme from Escherichia coli strain BL21 (DE3) by nickel affinity chromatography, anion exchange chromatography, and gel filtration	Pseudomonas syringae

Substrates and Products (Substrate)

Substrates	Comment Substrates	Organism	Products	Comment (Products)	Rev.	Reac.
hydroxymethylbilane	-	Pseudomonas syringae	uroporphyrinogen III + H2O	-	?
hydroxymethylbilane	-	Pseudomonas syringae DC3000	uroporphyrinogen III + H2O	-	?

Synonyms

Synonyms	Comment	Organism
U3S	-	Pseudomonas syringae
Uroporphyrinogen III synthase	-	Pseudomonas syringae

Temperature Optimum [°C]

Temperature Optimum [°C]	Temperature Optimum Maximum [°C]	Comment	Organism
37	-	assay at	Pseudomonas syringae

pH Optimum

pH Optimum Minimum	pH Optimum Maximum	Comment	Organism
8.2	-	assay at	Pseudomonas syringae

General Information

General Information	Comment	Organism
metabolism	U3S is one of the key enzymes in the biosynthesis of tetrapyrrole compounds	Pseudomonas syringae
physiological function	U3S catalyzes the cyclization of the linear hydroxymethylbilane to uroporphyrinogen III. Arg219 is the key residue important for the catalytic activity of psU3S, Tyr162 is important for activity	Pseudomonas syringae