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Literature summary extracted from

  • Polo, L.M.; Gil-Ortiz, F.; Cantin, A.; Rubio, V.
    New insight into the transcarbamylase family: the structure of putrescine transcarbamylase, a key catalyst for fermentative utilization of agmatine (2012), PLoS ONE, 7, e31528.
    View publication on PubMedView publication on EuropePMC

Cloned(Commentary)

EC Number Cloned (Comment) Organism
2.1.3.3 enzyme protomer structure, overview Enterococcus faecalis
2.1.3.6 PTC protomer structure, overview Enterococcus faecalis

Crystallization (Commentary)

EC Number Crystallization (Comment) Organism
2.1.3.3 purified full-length enzyme or in complex with inhibitors N-(phosphonoacetyl)-putrescine or N-(phosphonoacetyl)-L-ornithine, and trunacted enzyme in complex with N-(phosphonoacetyl)-L-ornithine, hanging drop vapor diffusion technique, mixing of 0.001 ml of 10 mg/ml protein in 50 mM Tris-HCl, pH 7.5 containing 0.43 mM ligand with 0.001 ml of crystallization solution composed of 125 mM (NH4)2SO4, 17% PEG 3350, 0.1 M Bis-Tris, pH 5.5, 21°C, X-ray diffraction structure determination and analysis at 2.5 A, 2.0 A, and 1.59 A resolution, respectively, modeling Enterococcus faecalis
2.1.3.6 purified full-length PTC or in complex with inhibitors N-(phosphonoacetyl)-putrescine or N-(phosphonoacetyl)-L-ornithine, and trunacted PTC in complex with N-(phosphonoacetyl)-L-ornithine, hanging drop vapor diffusion technique, mixing of 0.001 ml of 10 mg/ml protein in 50 mM Tris-HCl, pH 7.5 containing 0.43 mM ligand with 0.001 ml of crystallization solution composed of 125 mM (NH4)2SO4, 17% PEG 3350, 0.1 M Bis-Tris, pH 5.5, 21°C, X-ray diffraction structure determination and analysis at 2.5 A, 2.0 A, and 1.59 A resolution, respectively, modeling Enterococcus faecalis

Protein Variants

EC Number Protein Variants Comment Organism
2.1.3.3 additional information confirmation of decreased stability of the trimer of the enzyme lacking the C-terminal helix by deleting this helix Enterococcus faecalis
2.1.3.3 R54G site-directed mutagenesis, inactive mutant, not exhibiting any PTC or OTC activity Enterococcus faecalis
2.1.3.3 Y230V/G231S/L232M/Y233G engineering of the 230-loop of the enzyme, by replacing the sequence 230YGLY233 of the putrescine signature by its OTC counterpart VSMG, favors the use of ornithine and impairs that of putrescine Enterococcus faecalis
2.1.3.6 additional information confirmation of decreased stability of the trimer of PTC lacking the C-terminal helix by deleting this helix Enterococcus faecalis
2.1.3.6 R54G site-directed mutagenesis, inactive mutant, not exhibiting any PTC or OTC activity Enterococcus faecalis
2.1.3.6 Y230V/G231S/L232M/Y233G engineering of the 230-loop of PTC, by replacing the sequence 230YGLY233 of the putrescine signature by its OTC counterpart VSMG, favors the use of ornithine and impairs that of putrescine Enterococcus faecalis

Inhibitors

EC Number Inhibitors Comment Organism Structure
2.1.3.3 delta-N-(phosphonoacetyl)-L-ornithine bisubstrate analogue inhibitor Enterococcus faecalis
2.1.3.6 N-(phosphonoacetyl)-putrescine bisubstrate analogue inhibitor Enterococcus faecalis

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
2.1.3.3 32
-
 L-ornithine recombinant enzyme mutant Y230V/G231S/L232M/Y233G, pH 8.5, 37°C Enterococcus faecalis
2.1.3.3 36.4
-
 L-ornithine recombinant wild-type enzyme, pH 8.5, 37°C Enterococcus faecalis
2.1.3.6 1.14
-
 putrescine recombinant wild-type PTC, pH 8.5, 37°C Enterococcus faecalis
2.1.3.6 32.9
-
 putrescine recombinant PTC mutant Y230V/G231S/L232M/Y233G, pH 8.5, 37°C Enterococcus faecalis

Molecular Weight [Da]

EC Number Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
2.1.3.3 120300
-
 trimeric enzyme, gel filtration Enterococcus faecalis
2.1.3.3 230000
-
 hexameric enzyme, gel filtration Enterococcus faecalis
2.1.3.6 120300
-
 trimeric PTC, gel filtration Enterococcus faecalis
2.1.3.6 230000
-
 hexameric PTC, gel filtration Enterococcus faecalis

Natural Substrates/ Products (Substrates)

EC Number Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
2.1.3.3 carbamoyl phosphate + L-ornithine Enterococcus faecalis
-
 L-citrulline + phosphate
-
 r
2.1.3.6 carbamoyl phosphate + putrescine Enterococcus faecalis
-
 phosphate + N-carbamoylputrescine
-
 r

Organism

EC Number Organism UniProt Comment Textmining
2.1.3.3 Enterococcus faecalis Q837U7
-
-
2.1.3.6 Enterococcus faecalis Q837U7
-
-

Specific Activity [micromol/min/mg]

EC Number Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
2.1.3.3 24
-
 recombinant wild-type enzyme, substrate L-ornithine, pH 8.5, 37°C Enterococcus faecalis
2.1.3.3 82
-
 recombinant enzyme mutant Y230V/G231S/L232M/Y233G, substrate L-ornithine, pH 8.5, 37°C Enterococcus faecalis
2.1.3.6 1.13
-
 recombinant PTC mutant Y230V/G231S/L232M/Y233G, substrate putrescine, pH 8.5, 37°C Enterococcus faecalis
2.1.3.6 813
-
 recombinant wild-type PTC, substrate putrescine, pH 8.5, 37°C Enterococcus faecalis

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
2.1.3.3 carbamoyl phosphate + L-ornithine
-
 Enterococcus faecalis L-citrulline + phosphate
-
 r
2.1.3.3 additional information in addition to using putrescine, see EC 2.1.3.6, the enzyme can utilize L-ornithine as a poor substrate. Differences between the respective 230 and SMG loops of putrescine transcarbamoylase PTC and OTC appear to account for the differential preference of these enzymes for putrescine and ornithine, active center and the discrimination mechanism between putrescine and ornithine, overview Enterococcus faecalis ?
-
 ?
2.1.3.6 carbamoyl phosphate + putrescine
-
 Enterococcus faecalis phosphate + N-carbamoylputrescine
-
 r
2.1.3.6 additional information in addition to using putrescine, the enzyme can utilize L-ornithine as a poor substrate, see EC 2.1.3.3. Differences between the respective 230 and SMG loops of PTC and OTC appear to account for the differential preference of these enzymes for putrescine and ornithine, active center and the discrimination mechanism between putrescine and ornithine, overview Enterococcus faecalis ?
-
 ?

Subunits

EC Number Subunits Comment Organism
2.1.3.3 hexamer two trimer structure Enterococcus faecalis
2.1.3.3 More presence or absence of supratrimeric oligomerization, structure comparison and analysis, overview Enterococcus faecalis
2.1.3.6 hexamer two trimer structure Enterococcus faecalis
2.1.3.6 More presence or absence of supratrimeric oligomerization, structure comparison and analysis, overview Enterococcus faecalis

Synonyms

EC Number Synonyms Comment Organism
2.1.3.3 ornithine transcarbamylase
-
 Enterococcus faecalis
2.1.3.6 putrescine transcarbamylase
-
 Enterococcus faecalis

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
2.1.3.3 37
-
 assay at Enterococcus faecalis
2.1.3.6 37
-
 assay at Enterococcus faecalis

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
2.1.3.3 8.5
-
 assay at Enterococcus faecalis
2.1.3.6 8.5
-
 assay at Enterococcus faecalis

General Information

EC Number General Information Comment Organism
2.1.3.3 additional information sequence 230YGLY233 is the putrescine signature sequence Enterococcus faecalis
2.1.3.3 physiological function the OTC activity of the enzyme is involved in arginine biosynthesis Enterococcus faecalis
2.1.3.6 additional information sequence 230YGLY233 is the putrescine signature sequence Enterococcus faecalis
2.1.3.6 physiological function the enzyme generates carbamoyl phosphate for ATP production in the fermentative catabolism of agmatine Enterococcus faecalis