This enzyme is involved in the biosynthesis of factor 420, a redox-active cofactor, in methanogenic archaea and certain bacteria. The specific reaction catalysed in vivo is determined by the availability of substrate, which in turn is determined by the enzyme present in the organism - EC 2.7.7.68, 2-phospho-L-lactate guanylyltransferase, EC 2.7.7.105, phosphoenolpyruvate guanylyltransferase, or EC 2.7.7.106, 3-phospho-D-glycerate guanylyltransferase.
Specify your search results
The expected taxonomic range for this enzyme is: Bacteria, Archaea
This enzyme is involved in the biosynthesis of factor 420, a redox-active cofactor, in methanogenic archaea and certain bacteria. The specific reaction catalysed in vivo is determined by the availability of substrate, which in turn is determined by the enzyme present in the organism - EC 2.7.7.68, 2-phospho-L-lactate guanylyltransferase, EC 2.7.7.105, phosphoenolpyruvate guanylyltransferase, or EC 2.7.7.106, 3-phospho-D-glycerate guanylyltransferase.
catalysis does not appear to proceed via a covalent intermediate. Lactyl-2-diphospho-5'-guanosine is the preferred substrate. The substrate binding site of CofD is very specific for 7,8-didemethyl-8-hydroxy-5-deazariboflavin
to a lesser extent CofD also catalyzes a number of additional reactions that include the formation of 7,8-didemethyl-8-hydroxy-5-deazariboflavin 5'-phosphate, when the enzyme is incubated with 7,8-didemethyl-8-hydroxy-5-deazariboflavin and GDP, GTP, diphosphate, or tripolyphosphate, and the hydrolysis of F420-0 (F420 without glutamic acids) to 7,8-didemethyl-8-hydroxy-5-deazariboflavin
to a lesser extent CofD also catalyzes a number of additional reactions that include the formation of 7,8-didemethyl-8-hydroxy-5-deazariboflavin 5'-phosphate, when the enzyme is incubated with 7,8-didemethyl-8-hydroxy-5-deazariboflavin and GDP, GTP, diphosphate, or tripolyphosphate, and the hydrolysis of F420-0 (F420 without glutamic acids) to 7,8-didemethyl-8-hydroxy-5-deazariboflavin
isolated CofD enzyme is not activated by the addition of 0-4 mM MgCl2. The inactivation by EDTA can be completely reversed by the addition of excess Mg2+ but not by the addition of Zn2+ or Mn2+
CofD activity is measured in a coupled assay with lactylphosphate guanylyltransferase (CofC, MJ1117), which produces lactyl-2-diphospho-5'-guanosine from 2-phospho-L-lactate and GTP. The reaction mixture contains 50 mM TES (pH 7.5), 0.1 mM GTP, 10 mM 2-phospho-L-lactate, 0.4 mM 7,8-didemethyl-8-hydroxy-5-deazariboflavin, 2 mM MnCl2, 0.01 mg of CofC, and 0.00040.0006 mg of CofD. The enzyme system is inhibited by high concentrations of GMP. Therefore, 2 mM phosphoenolpyruvate and 1 unit of pyruvate kinase are also included in the reaction mixture to regenerate GTP from GMP
CofD activity is measured in a coupled assay with lactylphosphate guanylyltransferase (CofC, MJ1117), which produces lactyl-2-diphospho-5'-guanosine from 2-phospho-L-lactate and GTP. The reaction mixture contains 50 mM TES (pH 7.5), 0.1 mM GTP, 10 mM 2-phospho-L-lactate, 0.4 mM 7,8-didemethyl-8-hydroxy-5-deazariboflavin, 2 mM MnCl2, 0.01 mg of CofC, and 0.00040.0006 mg of CofD. The enzyme system is inhibited by high concentrations of GMP. Therefore, 2 mM phosphoenolpyruvate and 1 unit of pyruvate kinase are also included in the reaction mixture to regenerate GTP from GMP
Please wait a moment until the data is sorted. This message will disappear when the data is sorted.
CRYSTALLIZATION (Commentary)
ORGANISM
UNIPROT
LITERATURE
crystal structures of the CofD enzyme, in its free form at 3.1 A resolution, in a ternary complex with 7,8-didemethyl-8-hydroxy-5-deazaflavin and phosphate at 2.5 A resolution, and in a ternary complex with 7,8-didemethyl-8-hydroxy-5-deazaflavin and GDP at 3.0 A resolution
2.7.8.28 2-phospho-L-lactate transferase
more
top
