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Literature summary for 2.1.3.6 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)

Cloned (Comment) Organism
PTC protomer structure, overview Enterococcus faecalis

Crystallization (Commentary)

Crystallization (Comment) Organism
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

Protein Variants Comment Organism
additional information confirmation of decreased stability of the trimer of PTC lacking the C-terminal helix by deleting this helix Enterococcus faecalis
R54G site-directed mutagenesis, inactive mutant, not exhibiting any PTC or OTC activity Enterococcus faecalis
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

Inhibitors Comment Organism Structure
N-(phosphonoacetyl)-putrescine bisubstrate analogue inhibitor Enterococcus faecalis

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
1.14
-
putrescine recombinant wild-type PTC, pH 8.5, 37°C Enterococcus faecalis
32.9
-
putrescine recombinant PTC mutant Y230V/G231S/L232M/Y233G, pH 8.5, 37°C Enterococcus faecalis

Molecular Weight [Da]

Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
120300
-
trimeric PTC, gel filtration Enterococcus faecalis
230000
-
hexameric PTC, gel filtration Enterococcus faecalis

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
carbamoyl phosphate + putrescine Enterococcus faecalis
-
phosphate + N-carbamoylputrescine
-
r

Organism

Organism UniProt Comment Textmining
Enterococcus faecalis Q837U7
-
-

Specific Activity [micromol/min/mg]

Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
1.13
-
recombinant PTC mutant Y230V/G231S/L232M/Y233G, substrate putrescine, pH 8.5, 37°C Enterococcus faecalis
813
-
recombinant wild-type PTC, substrate putrescine, pH 8.5, 37°C Enterococcus faecalis

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
carbamoyl phosphate + putrescine
-
Enterococcus faecalis phosphate + N-carbamoylputrescine
-
r
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

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

Synonyms

Synonyms Comment Organism
putrescine transcarbamylase
-
Enterococcus faecalis

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
37
-
assay at Enterococcus faecalis

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
8.5
-
assay at Enterococcus faecalis

General Information

General Information Comment Organism
additional information sequence 230YGLY233 is the putrescine signature sequence Enterococcus faecalis
physiological function the enzyme generates carbamoyl phosphate for ATP production in the fermentative catabolism of agmatine Enterococcus faecalis