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Literature summary for 6.3.4.2 extracted from

  • Chang, Y.F.; Martin, S.S.; Baldwin, E.P.; Carman, G.M.
    Phosphorylation of human CTP synthetase 1 by protein kinase C: identification of Ser(462) and Thr(455) as major sites of phosphorylation (2007), J. Biol. Chem., 282, 17613-17622.
    View publication on PubMedView publication on EuropePMC

Cloned(Commentary)

Cloned (Comment) Organism
protein is expressed as a His6-tagged wild type human CTP synthetase 1 fusion protein in Escherichia coli Homo sapiens
protein is expressed as a His6-tagged wild type human CTP synthetase 1 fusion protein in Saccharomyces cerevisiae Homo sapiens

Protein Variants

Protein Variants Comment Organism
S462A/T455A S462A and T455A mutations result in a decreased CTP synthetase 1 phosphorylation which appear to be much less than of the individual mutant enzymes S462A or T455A Homo sapiens

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.002
-
ATP
-
Homo sapiens

Metals/Ions

Metals/Ions Comment Organism Structure
Mg2+ assay with 10 mM MgCl2 Homo sapiens

Organism

Organism UniProt Comment Textmining
Homo sapiens
-
-
-

Posttranslational Modification

Posttranslational Modification Comment Organism
phosphoprotein CTP synthetase is phosphorylated by protein kinase C which leads to 95fold higher activity of CTP synthetase 1. Phosphopeptide mapping and phosphoamino acid analyses shows that CTP synthetase 1 is phosphorylated on multiple serine and threonine residues. It is indicated that protein kinase C phosphorylation at Ser462 stimulates human CTP synthetase 1 activity, whereas phosphorylation at Thr455 inhibits activity Homo sapiens

Purification (Commentary)

Purification (Comment) Organism
His6-tagged wild type and mutant human CTP synthetase 1 enzymes are expressed and purified from yeast cell extracts with Ni2+-NTA resin Homo sapiens
His6-tagged wild type human CTP synthetase 1 is expressed and purified from Escherichia coli by Ni2+-NTA chromatography and with PorosHQ-ion-exchange chromatography Homo sapiens

Specific Activity [micromol/min/mg]

Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
additional information
-
S462A mutation results in 61%-reduced CTP synthetase 1 phosphorylation. The Saccharomyces cerevisiae-expressed and purified S462A mutant enzyme exhibits a 2fold reduction in CTP synthetase 1 activity, whereas the purified T455A mutant enzyme exhibited a 2fold elevation in CTP synthetase 1 activity, implying that that protein kinase C phosphorylation at Ser462 stimulates human CTP synthetase 1 activity, whereas phosphorylation at Thr455 inhibits activity Homo sapiens
additional information
-
T455A mutation results in 58%-reduced CTP synthetase 1 phosphorylation. The Saccharomyces cerevisiae-expressed and purified S462A mutant enzyme exhibits a 2fold reduction in CTP synthetase 1 activity, whereas the purified T455A mutant enzyme exhibited a 2fold elevation in CTP synthetase 1 activity, implying that that protein kinase C phosphorylation at Ser462 stimulates human CTP synthetase 1 activity, whereas phosphorylation at Thr455 inhibits activity Homo sapiens
0.0025
-
specific activity of Escherichia coli expressed enzyme, after a 20 min incubation with protein kinase C, the activity of the CTP synthetase was stimulated 95fold Homo sapiens

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
ATP + UTP + Gln + H2O
-
Homo sapiens ADP + CTP + Glu + phosphate
-
?

Synonyms

Synonyms Comment Organism
CTP synthetase 1
-
Homo sapiens

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
8
-
assay at Homo sapiens