Literature summary for 3.5.1.54 extracted from

Fan, C.; Li, Z.; Yin, H.; Xiang, S.
Structure and function of allophanate hydrolase (2013), J. Biol. Chem., 288, 21422-21432.

Cloned(Commentary)

Cloned (Comment)	Organism
recombinant expression of wild-type and mutant His-tagged SeMet-substituted enzymes in Escherichia coli strain BL21 Star(DE3)	Kluyveromyces lactis

Crystallization (Commentary)

Crystallization (Comment)	Organism
purified recombinant wild-type and mutant His-tagged SeMet-substituted enzymes, sitting drop vapour diffusion method, mixing of 10 mg/ml protein in 20 mM Tris/HCl, pH 7.5, 200 mM NaCl, 2-10 mM DTT, with reservoir containing 16% PEG 8000, 20% glycerol, and 0.04 M potassium phosphate, and 100 mM sodium/potassium tartrate, 20°C, X-ray diffraction structure determination and analysis at 2.5-2.6 A resolution, molecular replacement	Kluyveromyces lactis

Crystallization (Comment)

Organism

purified recombinant wild-type and mutant His-tagged SeMet-substituted enzymes, sitting drop vapour diffusion method, mixing of 10 mg/ml protein in 20 mM Tris/HCl, pH 7.5, 200 mM NaCl, 2-10 mM DTT, with reservoir containing 16% PEG 8000, 20% glycerol, and 0.04 M potassium phosphate, and 100 mM sodium/potassium tartrate, 20°C, X-ray diffraction structure determination and analysis at 2.5-2.6 A resolution, molecular replacement

Kluyveromyces lactis

Protein Variants

Protein Variants	Comment	Organism
G559E/G572E	site-directed mutagenesis, crystal structure analysis, the mutant shows 14fold increaed Km for allophanate, and reduced substrate binding at the N-domain active site, but is catalytically active	Kluyveromyces lactis
additional information	generation of isolated domains	Kluyveromyces lactis
S177A	site-directed mutagenesis, crystal structure analysis	Kluyveromyces lactis

Organism

Organism	UniProt	Comment	Textmining
Kluyveromyces lactis	Q6CP22	-	-

Purification (Commentary)

Purification (Comment)	Organism
recombinant wild-type and mutant His-tagged SeMet-substituted enzymes from Escherichia coli strain BL21 Star(DE3) by nickel affinity chromatography and gel filtration	Kluyveromyces lactis

Reaction

Reaction	Comment	Organism	Reaction ID
urea-1-carboxylate + H2O = 2 CO2 + 2 NH3	two-step reaction catalytic mechanism of the C-domain, overview. The C-domain probably catalyzes a distinct form of decarboxylation reaction	Kluyveromyces lactis	a

Subunits

Subunits	Comment	Organism
dimer	enzyme domain architecture, overview. Both the N- and the C-domains require dimerization for their optimal activities	Kluyveromyces lactis

General Information

General Information	Comment	Organism
additional information	the enzyme's N and C domains catalyze sequential reactions, overview	Kluyveromyces lactis
physiological function	allophanate hydrolase is essential for urea utilization. The enzyme also has important functions in the eukaryotic pyrimidine nucleic acid precursor degradation pathway, the yeast-hypha transition that several pathogens utilize to escape the host defense, and an s-triazine herbicide degradation pathway in soil bacteria	Kluyveromyces lactis