Literature summary extracted from

Cheng, J.H.; Huang, Y.H.; Lin, J.J.; Huang, C.Y.
Crystal structures of monometallic dihydropyrimidinase and the human dihydroorotase domain K1556A mutant reveal no lysine carbamylation within the active site (2018), Biochem. Biophys. Res. Commun., 505, 439-444 .

Cloned(Commentary)

EC Number	Cloned (Comment)	Organism
3.5.2.2	recombinant expression of His-tagged enzyme	Pseudomonas aeruginosa
3.5.2.3	recombinant expression of the mutant His-tagged enzyme	Homo sapiens

Crystallization (Commentary)

EC Number	Crystallization (Comment)	Organism
3.5.2.2	mono-Zn PaDHPase, hanging drop vapor diffusion method, mixing of 20 mg/ml protein in 20 mM HEPES and 100 mM NaCl, pH 7.0, with reservoir solution containing 25% PEG 4000, 100 mM Tris-HCl, 200 mM calcium chloride, pH 8.5, at room temperature, X-ray diffraction structure determination and analysis at 2.23 A resolution	Pseudomonas aeruginosa
3.5.2.3	human DHOase domain K1556A mutant, hanging drop vapor diffusion method, mixing of 20 mg/ml protein in 20 mM HEPES and 100 mM NaCl, pH 7.0, with reservoir solution containing 2 M sodium chloride, 100 mM MES, 200 mM sodium acetate, pH 6.5, at room temperature, X-ray diffraction structure determination and analysis at 2.77 A resolution	Homo sapiens

Protein Variants

EC Number	Protein Variants	Comment	Organism
3.5.2.3	K1556A	site-directed mutagenesis, the mutant shows no lysine carbamylation within the active site	Homo sapiens

Inhibitors

EC Number	Inhibitors	Comment	Organism	Structure
3.5.2.2	additional information	EDTA alone is a poor chelator for removal of metal in PaDHPase	Pseudomonas aeruginosa

Metals/Ions

EC Number	Metals/Ions	Comment	Organism	Structure
3.5.2.2	additional information	EDTA alone is a poor chelator for removal of metal in PaDHPase	Pseudomonas aeruginosa
3.5.2.2	Zn2+	di-zinc metalloenzyme, one metal coordination is not sufficient to fix CO2 to the Lys in bacterial DHPase. Preparation and characterization of mono-Zn DHPase from Pseudomonas aeruginosa revealing no catalytic activity of mono-Zn PaDHPase can be detected, Lys150 is no longer carbamylated, mass spectrometric analysis. Posttranslational carbamylated Lys is not required for Znalpha binding in PaDHPase. Znalpha is coordinated by residues H59, H61, K150, D316, H183, and H239 in mono-Zn PaDHPase. In di-zinf PaDHPase, zinc alpha is coordinated by residues H59, H61, Kcx150, and D316, while zinc beta is coordinated by H183, H239, K150, and zinc alpha	Pseudomonas aeruginosa
3.5.2.2	Zn2+	zinc metalloenzyme, one zinc ion is sufficient to stabilize Lys carbamylation in Tetraodon nigroviridis dihydropyrimidinase (DHPase). Lys155 is carbamylated, structure, overview	Tetraodon nigroviridis
3.5.2.3	Zn2+	mono-zinc metalloenzyme, the active site of the huDHOase K1556A mutant contains one metal ion. Posttranslational carbamylated Lys is not required for Znalpha binding in huDHOase	Homo sapiens

Natural Substrates/ Products (Substrates)

EC Number	Natural Substrates	Organism	Comment (Nat. Sub.)	Natural Products	Comment (Nat. Pro.)	Rev.	Reac.
3.5.2.3	(S)-dihydroorotate + H2O	Homo sapiens	-	N-carbamoyl-L-aspartate	-	?

Organism

EC Number	Organism	UniProt	Comment	Textmining
3.5.2.2	Pseudomonas aeruginosa	Q9I676	-	-
3.5.2.2	Pseudomonas aeruginosa 1C	Q9I676	-	-
3.5.2.2	Pseudomonas aeruginosa ATCC 15692	Q9I676	-	-
3.5.2.2	Pseudomonas aeruginosa CIP 104116	Q9I676	-	-
3.5.2.2	Pseudomonas aeruginosa DSM 22644	Q9I676	-	-
3.5.2.2	Pseudomonas aeruginosa JCM 14847	Q9I676	-	-
3.5.2.2	Pseudomonas aeruginosa LMG 12228	Q9I676	-	-
3.5.2.2	Pseudomonas aeruginosa PRS 101	Q9I676	-	-
3.5.2.2	Tetraodon nigroviridis	-	-	-
3.5.2.3	Homo sapiens	P27708	-	-

Purification (Commentary)

EC Number	Purification (Comment)	Organism
3.5.2.2	recombinant His-tagged enzyme by nickel affinity chromatography and dialysis to over 97% purity	Pseudomonas aeruginosa
3.5.2.3	recombinant mutant His-tagged enzyme by nickel affinity chromatography and dialysis	Homo sapiens

Substrates and Products (Substrate)

EC Number	Substrates	Comment Substrates	Organism	Products	Comment (Products)	Rev.	Reac.
3.5.2.2	additional information	one zinc ion is sufficient to stabilize Lys carbamylation in Tetraodon nigroviridis dihydropyrimidinase (DHPase)	Pseudomonas aeruginosa	?	-	-
3.5.2.2	additional information	one zinc ion is sufficient to stabilize Lys carbamylation in Tetraodon nigroviridis dihydropyrimidinase (DHPase)	Tetraodon nigroviridis	?	-	-
3.5.2.2	additional information	one zinc ion is sufficient to stabilize Lys carbamylation in Tetraodon nigroviridis dihydropyrimidinase (DHPase)	Pseudomonas aeruginosa ATCC 15692	?	-	-
3.5.2.2	additional information	one zinc ion is sufficient to stabilize Lys carbamylation in Tetraodon nigroviridis dihydropyrimidinase (DHPase)	Pseudomonas aeruginosa 1C	?	-	-
3.5.2.2	additional information	one zinc ion is sufficient to stabilize Lys carbamylation in Tetraodon nigroviridis dihydropyrimidinase (DHPase)	Pseudomonas aeruginosa PRS 101	?	-	-
3.5.2.2	additional information	one zinc ion is sufficient to stabilize Lys carbamylation in Tetraodon nigroviridis dihydropyrimidinase (DHPase)	Pseudomonas aeruginosa DSM 22644	?	-	-
3.5.2.2	additional information	one zinc ion is sufficient to stabilize Lys carbamylation in Tetraodon nigroviridis dihydropyrimidinase (DHPase)	Pseudomonas aeruginosa CIP 104116	?	-	-
3.5.2.2	additional information	one zinc ion is sufficient to stabilize Lys carbamylation in Tetraodon nigroviridis dihydropyrimidinase (DHPase)	Pseudomonas aeruginosa LMG 12228	?	-	-
3.5.2.2	additional information	one zinc ion is sufficient to stabilize Lys carbamylation in Tetraodon nigroviridis dihydropyrimidinase (DHPase)	Pseudomonas aeruginosa JCM 14847	?	-	-
3.5.2.3	(S)-dihydroorotate + H2O	-	Homo sapiens	N-carbamoyl-L-aspartate	-	?

Synonyms

EC Number	Synonyms	Comment	Organism
3.5.2.2	DHPase	-	Pseudomonas aeruginosa
3.5.2.2	DHPase	-	Tetraodon nigroviridis
3.5.2.2	PaDHPase	-	Pseudomonas aeruginosa
3.5.2.3	DHOase	-	Homo sapiens
3.5.2.3	huDHOase	-	Homo sapiens
3.5.2.3	human DHOase domain	-	Homo sapiens
3.5.2.3	More	cf. EC 6.3.5.5	Homo sapiens

General Information

EC Number	General Information	Comment	Organism
3.5.2.2	evolution	dihydropyrimidinase (DHPase) is a member of the cyclic amidohydrolase family, which also includes allantoinase, dihydroorotase (DHOase), hydantoinase, and imidase. Almost all of these zinc metalloenzymes possess a binuclear metal center in which two metal ions are bridged by a post-translational carbamylated Lys	Pseudomonas aeruginosa
3.5.2.2	evolution	dihydropyrimidinase (DHPase) is a member of the cyclic amidohydrolase family, which also includes allantoinase, dihydroorotase (DHOase), hydantoinase, and imidase. Almost all of these zinc metalloenzymes possess a binuclear metal center in which two metal ions are bridged by a post-translational carbamylated Lys	Tetraodon nigroviridis
3.5.2.2	additional information	structure-activity relationship analysis	Pseudomonas aeruginosa
3.5.2.3	evolution	dihydroorotase (DHOase) is a member of the cyclic amidohydrolase family, which also includes allantoinase, hydantoinase, dihydropyrimidinase (DHPase), and imidase. Almost all of these zinc metalloenzymes possess a binuclear metal center in which two metal ions are bridged by a post-translational carbamylated Lys	Homo sapiens
3.5.2.3	additional information	structure-activity relationship analysis	Homo sapiens

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Release 2023.1 (January 2023)