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Literature summary for 6.3.4.2 extracted from

  • Braun, O.; Knipp, M.; Chesnov, S.; Vasak, M.
    Specific reactions of S-nitrosothiols with cysteine hydrolases: A comparative study between dimethylargininase-1 and CTP synthetase (2007), Protein Sci., 16, 1522-1534.
    View publication on PubMedView publication on EuropePMC
Search for more literature for 6.3.4.2

Inhibitors

Inhibitors Comment Organism Structure
additional information incubation of EcCTPS modified by CysNO and HcyNO with 5 mM DTT for 30 min at 37°C reveals that 88% and 97%, respectively, of the original activity can be recovered; it is shown that in the presence of 1 mM Gln, S-nitroso-L-cysteine reduces the enzymatic activity by 88% and by 32% in the presence of 10 mM Gln. Similar studies with S-nitroso-L-homocysteine result in reduction of the activity by 43% and 19%, respectively. The results suggest that the substrate Gln competitively protects the active site of EcCTPS from the modification with S-nitroso-L-cysteine and S-nitroso-L-homocysteine.; no inhibition by S-nitrosoglutathione presumably due to its inability to enter the actve site of the enzyme Escherichia coli
S-nitroso-L-cysteine specific irreversible inhibitor,inhibits the activity by 94% Escherichia coli
S-nitroso-L-homocysteine specific irreversible inhibitor, inhibits the activity by 90% Escherichia coli

Metals/Ions

Metals/Ions Comment Organism Structure
Mg2+ assay with 10 mM Mg2+ Escherichia coli

Molecular Weight [Da]

Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
60240
-
 mass spectroscopy, processed N-terminal Met1 Escherichia coli
60380
-
 mass spectroscopy, unprocessed N-terminal Met1 Escherichia coli

Organism

Organism UniProt Comment Textmining
Escherichia coli P0A7E5
-
-

Purification (Commentary)

Purification (Comment) Organism
enzyme is purified by gel filtration on a HiLoad 26/60 Superdex 200 column. Active fractions are pooled and applied to a Mono Q HR 5/5 column. Final purifications step is performed on a HiLoad 26/60 Superdex 75 column. The isolated EcCTPS is found >90% pure as judged by SDS-PAGE and ESI-MS. Escherichia coli

Synonyms

Synonyms Comment Organism
CTP synthetase
-
 Escherichia coli
EcCTPS
-
 Escherichia coli

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
37
-
 assay at Escherichia coli

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
7.3
-
 assay at Escherichia coli

Ki Value [mM]

Ki Value [mM] Ki Value maximum [mM] Inhibitor Comment Organism Structure
additional information
-
 additional information no saturation kinetic is obtained with S-nitroso-L-homocysteine Escherichia coli
0.027
-
 S-nitroso-L-cysteine Kinact: 0.48/min Escherichia coli