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Literature summary 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

Activating Compound

EC Number Activating Compound Comment Organism Structure
2.7.7.2 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
2.7.7.2 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)

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

Protein Variants

EC Number Protein Variants Comment Organism
2.7.7.2 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
2.7.7.2 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

EC Number Inhibitors Comment Organism Structure
2.7.7.2 ATP
-
 Methanocaldococcus jannaschii

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
2.7.7.2 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
2.7.7.2 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
2.7.7.2 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

EC Number Metals/Ions Comment Organism Structure
2.7.7.2 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
2.7.7.2 Fe2+ the enzyme contains 0.2 mol of iron per protomer Methanocaldococcus jannaschii
2.7.7.2 Mg2+ low stimulation of activity Methanocaldococcus jannaschii
2.7.7.2 Mg2+ the enzyme contains 8.4 mol of magnesium per protomer Methanocaldococcus jannaschii
2.7.7.2 Mn2+ the activity with Mn2+ is 4times lower than that with Co2+ Methanocaldococcus jannaschii
2.7.7.2 additional information the enzyme activity is not stimulated by Zn2+ and Fe2+ Methanocaldococcus jannaschii
2.7.7.2 Ni2+ weak stimulation of activity Methanocaldococcus jannaschii

Molecular Weight [Da]

EC Number Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
2.7.7.2 17000
-
 2 * 17000, SDS-PAGE Methanocaldococcus jannaschii
2.7.7.2 29000
-
 gel filtration Methanocaldococcus jannaschii

Organism

EC Number Organism UniProt Comment Textmining
2.7.7.2 Methanocaldococcus jannaschii Q58579
-
-

Oxidation Stability

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

Purification (Commentary)

EC Number Purification (Comment) Organism
2.7.7.2 Mono Q column chromatography Methanocaldococcus jannaschii

Specific Activity [micromol/min/mg]

EC Number Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
2.7.7.2 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
2.7.7.2 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
2.7.7.2 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)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
2.7.7.2 ATP + FMN the enzyme does not catalyze the reverse reaction to produce FMN and ATP from FAD and diphosphate Methanocaldococcus jannaschii diphosphate + FAD
-
 ir
2.7.7.2 CTP + FMN
-
 Methanocaldococcus jannaschii diphosphate + flavin cytidine dinucleotide
-
 ?
2.7.7.2 GTP + FMN weak specific activity Methanocaldococcus jannaschii diphosphate + flavin guanidine dinucleotide
-
 ?
2.7.7.2 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

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

Synonyms

EC Number Synonyms Comment Organism
2.7.7.2 MJ1179
-
 Methanocaldococcus jannaschii
2.7.7.2 RibL
-
 Methanocaldococcus jannaschii

Ki Value [mM]

EC Number Ki Value [mM] Ki Value maximum [mM] Inhibitor Comment Organism Structure
2.7.7.2 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]

EC Number kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
2.7.7.2 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
2.7.7.2 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
2.7.7.2 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