Cloned (Comment) | Organism |
---|---|
expression in Escherichia coli | Methanosarcina mazei |
Crystallization (Comment) | Organism |
---|---|
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 | Methanosarcina mazei |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
lactyl-2-diphospho-5'-guanosine + 7,8-didemethyl-8-hydroxy-5-deazariboflavin | Methanosarcina mazei | the reaction is the fourth step in the biosynthesis of coenzyme F420 | guanosine 5'-phosphate + coenzyme F420-0 | coenzyme F420-0 is coenzyme F420 without glutamic acid | ? |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Methanosarcina mazei | Q8PVT6 | - |
- |
Purification (Comment) | Organism |
---|---|
- |
Methanosarcina mazei |
Specific Activity Minimum [µmol/min/mg] | Specific Activity Maximum [µmol/min/mg] | Comment | Organism |
---|---|---|---|
additional information | - |
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 | Methanosarcina mazei |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
lactyl-2-diphospho-5'-guanosine + 7,8-didemethyl-8-hydroxy-5-deazariboflavin | the reaction is the fourth step in the biosynthesis of coenzyme F420 | Methanosarcina mazei | guanosine 5'-phosphate + coenzyme F420-0 | coenzyme F420-0 is coenzyme F420 without glutamic acid | ? |
Synonyms | Comment | Organism |
---|---|---|
2-phospho-L-lactate transferase | - |
Methanosarcina mazei |
CofD | - |
Methanosarcina mazei |