Cloned (Comment) | Organism |
---|---|
overexpression in Escherichia coli | Clostridioides difficile |
Crystallization (Comment) | Organism |
---|---|
crystals of the dehydratase are grown at 16°C by the vapor diffusion technique from a reservoir solution containing 18-29% (w/v) PEG 3350 and 100 mM BisTris (pH 6.5). The dehydratase is cocrystallized by incubation with 5 mM (R)-2-hydroxyisocaproate 10 min prior to crystallization | Clostridioides difficile |
Metals/Ions | Comment | Organism | Structure |
---|---|---|---|
[4Fe-4S] center | the crystal structure reveals that the heterodimeric protein contains two [4Fe-4S] clusters at a distance of 12 A, each coordinated by three cysteines and one terminal ligand. The cluster in the alpha-subunit is part of the active site. In the absence of substrate, a water/hydroxide ion acts as the fourth ligand. The substrate replaces this ligand and coordinates the cluster via the carbonyl-oxygen of the thioester group. The cluster in the beta-subunit has a terminal sulfhydryl/sulfido ligand and can act as a reservoir to protect the electron from unwanted side reactions via a recycling mechanism | Clostridioides difficile |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
(R)-2-hydroxy-4-methylpentanoyl-CoA | Clostridioides difficile | the enzyme is involved in fermentation of L-leucine | 4-methylpent-2-enoyl-CoA + H2O | - |
? |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Clostridioides difficile | Q5U923 and Q5U924 | hadC: Q5U923, hadB: Q5U924 | - |
Purification (Comment) | Organism |
---|---|
- |
Clostridioides difficile |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
(R)-2-hydroxy-4-methylpentanoyl-CoA | the enzyme is involved in fermentation of L-leucine | Clostridioides difficile | 4-methylpent-2-enoyl-CoA + H2O | - |
? | |
(R)-2-hydroxy-4-methylpentanoyl-CoA | the catalyzed reaction, an atypical alpha/beta-dehydration, depends on the reductive formation of ketyl radicals on the substrate generated by injection of a single electron from the ATP-dependent activator protein | Clostridioides difficile | 4-methylpent-2-enoyl-CoA + H2O | - |
? |
Subunits | Comment | Organism |
---|---|---|
heterodimer | - |
Clostridioides difficile |
Synonyms | Comment | Organism |
---|---|---|
HadBC | - |
Clostridioides difficile |