EC Number | Cloned (Comment) | Organism |
---|---|---|
1.17.7.4 | cloning from genomic DNA and expression as His-tagged protein in Escherichia coli strain XL-1 Blue | Escherichia coli |
EC Number | Crystallization (Comment) | Organism |
---|---|---|
1.17.7.4 | purified recombinant IspH, sitting drop vapour diffusion method at 20°C, 0.004 ml of 35 /ml protein in 50 mM Tris-HCl, pH 8.0, is mixed with 0.004 ml of reservoir solution containing 100 mM Bis-Tris-HCl, pH 6.5, 200 mM lithium sulfate and 25% PEG 3350, final pH is 6.8, crystal soaking in 2 M lithium sulfate, pH 6.5, before freezing, X-ray diffraction structure determination and analysis at 1.4 A resolution, molecular replacement, modelling | Escherichia coli |
EC Number | General Stability | Organism |
---|---|---|
1.17.7.4 | IspH is more stable in its closed conformation with a substrate molecule bound to the cluster, compared to its open conformation in solution | Escherichia coli |
EC Number | Metals/Ions | Comment | Organism | Structure |
---|---|---|---|---|
1.17.7.4 | Fe2+ | IspH contains a [4Fe-4S] cluster | Escherichia coli |
EC Number | Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|---|
1.17.7.4 | (E)-4-hydroxy-3-methylbut-2-en-1-yl diphosphate + NAD(P)H + H+ | Escherichia coli | in the nonmevalonate pathway, the terminal intermediate, 1-hydroxy-2-methyl-2-butenyl 4-diphosphate, i.e. HMBPP, is reductively converted into a mixture of isopentenyl diphosphate and dimethylallyl diphosphate | dimethylallyl diphosphate + NAD(P)+ + H2O | - |
r | |
1.17.7.4 | (E)-4-hydroxy-3-methylbut-2-en-1-yl diphosphate + NAD(P)H + H+ | Escherichia coli | in the nonmevalonate pathway, the terminal intermediate, 1-hydroxy-2-methyl-2-butenyl 4-diphosphate, i.e. HMBPP, is reductively converted into a mixture of isopentenyl diphosphate and dimethylallyl diphosphate | isopentenyl diphosphate + NAD(P)+ + H2O | - |
r |
EC Number | Organism | UniProt | Comment | Textmining |
---|---|---|---|---|
1.17.7.4 | Escherichia coli | - |
gene ispH, formerly lytB | - |
EC Number | Purification (Comment) | Organism |
---|---|---|
1.17.7.4 | recombinant His-tagged IspH from strain XL-1 Blue by nickel affinity and anion exchange chromatography followed by dialysis, over 99% purity | Escherichia coli |
EC Number | Reaction | Comment | Organism | Reaction ID |
---|---|---|---|---|
1.17.7.4 | dimethylallyl diphosphate + 2 oxidized ferredoxin [iron-sulfur] cluster + H2O = (E)-4-hydroxy-3-methylbut-2-en-1-yl diphosphate + 2 reduced ferredoxin [iron-sulfur] cluster + 2 H+ | reaction mechanism, detailed overview | Escherichia coli | |
1.17.7.4 | isopentenyl diphosphate + 2 oxidized ferredoxin [iron-sulfur] cluster + H2O = (E)-4-hydroxy-3-methylbut-2-en-1-yl diphosphate + 2 reduced ferredoxin [iron-sulfur] cluster + 2 H+ | reaction mechanism, detailed overview | Escherichia coli |
EC Number | Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|---|
1.17.7.4 | (E)-4-hydroxy-3-methylbut-2-en-1-yl diphosphate + NAD(P)H + H+ | in the nonmevalonate pathway, the terminal intermediate, 1-hydroxy-2-methyl-2-butenyl 4-diphosphate, i.e. HMBPP, is reductively converted into a mixture of isopentenyl diphosphate and dimethylallyl diphosphate | Escherichia coli | dimethylallyl diphosphate + NAD(P)+ + H2O | - |
r | |
1.17.7.4 | (E)-4-hydroxy-3-methylbut-2-en-1-yl diphosphate + NAD(P)H + H+ | formation of a mixture of isopentenyl diphosphate and dimethylallyl diphosphate | Escherichia coli | dimethylallyl diphosphate + NAD(P)+ + H2O | - |
r | |
1.17.7.4 | (E)-4-hydroxy-3-methylbut-2-en-1-yl diphosphate + NAD(P)H + H+ | in the nonmevalonate pathway, the terminal intermediate, 1-hydroxy-2-methyl-2-butenyl 4-diphosphate, i.e. HMBPP, is reductively converted into a mixture of isopentenyl diphosphate and dimethylallyl diphosphate | Escherichia coli | isopentenyl diphosphate + NAD(P)+ + H2O | - |
r | |
1.17.7.4 | (E)-4-hydroxy-3-methylbut-2-en-1-yl diphosphate + NAD(P)H + H+ | formation of a mixture of isopentenyl diphosphate and dimethylallyl diphosphate | Escherichia coli | isopentenyl diphosphate + NAD(P)+ + H2O | - |
r | |
1.17.7.4 | additional information | active site structure analysis of the IspH protein in complex with the substrate and the reaction products, or with diphosphate, overview. IspH is more stable in its closed conformation with a substrate molecule bound to the cluster, compared to its open conformation in solution | Escherichia coli | ? | - |
? |
EC Number | Subunits | Comment | Organism |
---|---|---|---|
1.17.7.4 | monomer | - |
Escherichia coli |
EC Number | Synonyms | Comment | Organism |
---|---|---|---|
1.17.7.4 | ispH | - |
Escherichia coli |
EC Number | Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|---|
1.17.7.4 | 30 | - |
assay at | Escherichia coli |
EC Number | pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|---|
1.17.7.4 | 7.5 | - |
assay at | Escherichia coli |