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Literature summary extracted from

  • Butryn, A.; Stoehr, G.; Linke-Winnebeck, C.; Hopfner, K.P.
    Serendipitous crystallization and structure determination of cyanase (CynS) from Serratia proteamaculans (2015), Acta Crystallogr. Sect. F, 71, 471-476 .
    View publication on PubMedView publication on EuropePMC

Cloned(Commentary)

EC Number Cloned (Comment) Organism
4.2.1.104 gene cynS, recombinant expression of N-terminally His6-tagged enzyme in Escherichia coli strain Rosetta (DE3) or Trichoplusia ni High Five cells Serratia proteamaculans

Crystallization (Commentary)

EC Number Crystallization (Comment) Organism
4.2.1.104 enzyme SpCynS as DNA-protein complex is crystallized as an impurity and its identity is determined using mass-spectrometric analysis. The crystals seems to have a ligand bound in the active site, glycerol molecules are bound at the entry to the active site of the enzyme. Mixing of 500 nl of 5.5 mg/ml protein-DNA complex in 20 mM MES potassium, pH 6.5, 60 mM KCl, 5 mM MgCl2, 2 mM DTT, with 500 nl of reservoir solution containing 0.1 M HEPES, pH 7.5, 2% v/v 2-propanol, 0.1 M sodium acetate, 18% w/v PEG 8000, and equilibration against 0.055 ml of reservoir solution, four months at 20°C, X-ray diffraction structure determination and analysis at 2.1 A resolution, the cryoprotecting solution consists of mother liquor containing 25% v/v glycerol, the crystals belong to space group P1, molecular replacement solution using the target DNA-protein complex or its components as a search model and the EcCynS crystal structure (PDB ID 1dw9) as a search model, modeling without ligand Serratia proteamaculans

Molecular Weight [Da]

EC Number Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
4.2.1.104 172000
-
 homodecamer, gel filtration Serratia proteamaculans
4.2.1.104 220000
-
 protein-DNA complex, gel filtration Serratia proteamaculans

Natural Substrates/ Products (Substrates)

EC Number Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
4.2.1.104 cyanate + HCO3- + 2 H+ Serratia proteamaculans
-
 NH3 + 2 CO2
-
 ?
4.2.1.104 cyanate + HCO3- + 2 H+ Serratia proteamaculans 568
-
 NH3 + 2 CO2
-
 ?

Organism

EC Number Organism UniProt Comment Textmining
4.2.1.104 Serratia proteamaculans A8GBZ7
-
-
4.2.1.104 Serratia proteamaculans 568 A8GBZ7
-
-

Purification (Commentary)

EC Number Purification (Comment) Organism
4.2.1.104 recombinant N-terminally His6-tagged enzyme from Escherichia coli strain Rosetta (DE3) or Trichoplusia ni High Five cells by nickel affinity chromatography, anion and/or cation ion exchange chromatography, and gel filtration to over 90% purity Serratia proteamaculans

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
4.2.1.104 cyanate + HCO3- + 2 H+
-
 Serratia proteamaculans NH3 + 2 CO2
-
 ?
4.2.1.104 cyanate + HCO3- + 2 H+
-
 Serratia proteamaculans 568 NH3 + 2 CO2
-
 ?

Subunits

EC Number Subunits Comment Organism
4.2.1.104 homodecamer
-
 Serratia proteamaculans
4.2.1.104 homodimer
-
 Serratia proteamaculans
4.2.1.104 More CynS protomers form dimers through intricate interactions of their C-terminal domains. Both protomers contribute two beta-strands each to one continuous antiparallel beta-sheet, and the two beta-strands of one protomer are packed against a long alpha-helix from the other protomers within the dimer. Like EcCynS, SpCynS assembles into a ring-like decamer with 5/2 symmetry Serratia proteamaculans

Synonyms

EC Number Synonyms Comment Organism
4.2.1.104 CynS
-
 Serratia proteamaculans
4.2.1.104 SpCynS
-
 Serratia proteamaculans
4.2.1.104 Spro_1533
-
 Serratia proteamaculans

General Information

EC Number General Information Comment Organism
4.2.1.104 additional information glycerol molecules bound at the entry to the active site of the enzyme during crystallization indicate conserved residues that might be important for the trafficking of substrates and products. The enzyme binds specifically to DNA Serratia proteamaculans
4.2.1.104 physiological function cyanate hydratase (CynS) catalyzes the decomposition of cyanate and bicarbonate into ammonia and carbon dioxide Serratia proteamaculans