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Literature summary for 2.7.7.2 extracted from

  • Mashhadi, Z.; Xu, H.; Grochowski, L.L.; White, R.H.
    Archaeal RibL: a new FAD synthetase that is air sensitive (2010), Biochemistry, 49, 8748-8755.
    View publication on PubMed
Search for more literature for 2.7.7.2

Activating Compound

Activating Compound Comment Organism Structure
ATP the enzyme reaches its maximal activity at an ATP concentration of about 1.4 mM. The enzyme activity decreases by 20% when the concentration of ATP is higher than the physiologically relevant concentration (about 5 mM) Methanocaldococcus jannaschii
CTP the activity of the enzyme reaches its maximal activity at a CTP concentration of about 1.4 mM. The maximal activity decreases by 68 and 95% when the concentration of CTP is 5.7 and 11.4 mM, respectively Methanocaldococcus jannaschii

Cloned(Commentary)

Cloned (Comment) Organism
expressed in Escherichia coli BL21-Codon Plus (DE3)-RIL cells Methanocaldococcus jannaschii

Protein Variants

Protein Variants Comment Organism
C126S the mutation does not reduce the protein's heat stability or solubility, the mutant contains less than 0.8 and less than 0.08 mol of Mg and Fe per protomer. In the presence of MgCl2, the mutant has activity about 2times higher than that of the wild type enzyme. The activity of the mutant in presence of Co2+ is very low Methanocaldococcus jannaschii
C143S the mutation does not reduce the protein's heat stability or solubility, the mutant contains less than 0.8 and less than 0.08 mol of Mg and Fe per protomer. In the presence of MgCl2, the mutant has activity approximately wild type activity. The activity of the mutant in presence of Co2+ is very low Methanocaldococcus jannaschii

Inhibitors

Inhibitors Comment Organism Structure
ATP
-
 Methanocaldococcus jannaschii

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.025
-
 ATP apparent value, in 35 mM TES (K+) buffer (pH 7.2) containing 14 mM dithiothreitol, 7 mM MgCl+, at 70°C Methanocaldococcus jannaschii
0.063
-
 FMN apparent value, in 35 mM TES (K+) buffer (pH 7.2) containing 14 mM dithiothreitol, 7 mM MgCl+, at 70°C Methanocaldococcus jannaschii
0.48
-
 CTP apparent value, in 35 mM TES (K+) buffer (pH 7.2) containing 14 mM dithiothreitol, 7 mM MgCl+, at 70°C Methanocaldococcus jannaschii

Metals/Ions

Metals/Ions Comment Organism Structure
Co2+ the enzyme requires divalent metals for activity, the best activity being observed with Co2+, where the activity is 4times greater than that with Mg2+ Methanocaldococcus jannaschii
Fe2+ the enzyme contains 0.2 mol of iron per protomer Methanocaldococcus jannaschii
Mg2+ low stimulation of activity Methanocaldococcus jannaschii
Mg2+ the enzyme contains 8.4 mol of magnesium per protomer Methanocaldococcus jannaschii
Mn2+ the activity with Mn2+ is 4times lower than that with Co2+ Methanocaldococcus jannaschii
additional information the enzyme activity is not stimulated by Zn2+ and Fe2+ Methanocaldococcus jannaschii
Ni2+ weak stimulation of activity Methanocaldococcus jannaschii

Molecular Weight [Da]

Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
17000
-
 2 * 17000, SDS-PAGE Methanocaldococcus jannaschii
29000
-
 gel filtration Methanocaldococcus jannaschii

Organism

Organism UniProt Comment Textmining
Methanocaldococcus jannaschii Q58579
-
-

Oxidation Stability

Oxidation Stability Organism
aerobically isolated enzyme is active only under reducing conditions (with 5.6 mM dithiothreitol) Methanocaldococcus jannaschii

Purification (Commentary)

Purification (Comment) Organism
Mono Q column chromatography Methanocaldococcus jannaschii

Specific Activity [micromol/min/mg]

Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
0.001
-
 using GTP as cosubstrate, in 35 mM TES (K+) buffer (pH 7.2) containing 14 mM dithiothreitol, 7 mM MgCl+, at 70°C Methanocaldococcus jannaschii
0.004
-
 using CTP as cosubstrate, in 35 mM TES (K+) buffer (pH 7.2) containing 14 mM dithiothreitol, 7 mM MgCl+, at 70°C Methanocaldococcus jannaschii
0.01
-
 using ATP as cosubstrate, in 35 mM TES (K+) buffer (pH 7.2) containing 14 mM dithiothreitol, 7 mM MgCl+, at 70°C Methanocaldococcus jannaschii

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
ATP + FMN the enzyme does not catalyze the reverse reaction to produce FMN and ATP from FAD and diphosphate Methanocaldococcus jannaschii diphosphate + FAD
-
 ir
CTP + FMN
-
 Methanocaldococcus jannaschii diphosphate + flavin cytidine dinucleotide
-
 ?
GTP + FMN weak specific activity Methanocaldococcus jannaschii diphosphate + flavin guanidine dinucleotide
-
 ?
additional information the enzyme does not function as a glycerol-3-phosphate cytidylyltransferase because it fails to catalyze the formation of glycerol cytidine dinucleotide when incubated with DL-glycerol 3-phosphate and CTP Methanocaldococcus jannaschii ?
-
 ?

Subunits

Subunits Comment Organism
homodimer 2 * 17000, SDS-PAGE Methanocaldococcus jannaschii

Synonyms

Synonyms Comment Organism
MJ1179
-
 Methanocaldococcus jannaschii
RibL
-
 Methanocaldococcus jannaschii

Ki Value [mM]

Ki Value [mM] Ki Value maximum [mM] Inhibitor Comment Organism Structure
31
-
 ATP in 35 mM TES (K+) buffer (pH 7.2) containing 14 mM dithiothreitol, 7 mM MgCl+, at 70°C Methanocaldococcus jannaschii

kcat/KM [mM/s]

kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
0.006
-
 CTP apparent value, in 35 mM TES (K+) buffer (pH 7.2) containing 14 mM dithiothreitol, 7 mM MgCl+, at 70°C Methanocaldococcus jannaschii
0.064
-
 FMN apparent value, in 35 mM TES (K+) buffer (pH 7.2) containing 14 mM dithiothreitol, 7 mM MgCl+, at 70°C Methanocaldococcus jannaschii
0.16
-
 ATP apparent value, in 35 mM TES (K+) buffer (pH 7.2) containing 14 mM dithiothreitol, 7 mM MgCl+, at 70°C Methanocaldococcus jannaschii