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Literature summary extracted from

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

Cloned(Commentary)

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

Crystallization (Commentary)

EC Number Crystallization (Comment) Organism
3.5.1.54 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

EC Number Protein Variants Comment Organism
3.5.1.54 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
3.5.1.54 additional information generation of isolated domains Kluyveromyces lactis
3.5.1.54 S177A site-directed mutagenesis, crystal structure analysis Kluyveromyces lactis

Organism

EC Number Organism UniProt Comment Textmining
3.5.1.54 Kluyveromyces lactis Q6CP22
-
-

Purification (Commentary)

EC Number Purification (Comment) Organism
3.5.1.54 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

EC Number Reaction Comment Organism Reaction ID
3.5.1.54 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

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

General Information

EC Number General Information Comment Organism
3.5.1.54 additional information the enzyme's N and C domains catalyze sequential reactions, overview Kluyveromyces lactis
3.5.1.54 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