Cloned (Comment) | Organism |
---|---|
expression of the enzyme with a thrombin-cleavable N-terminal His6 tag in Escherichia coli strain BL21(DE3) | Mycobacterium tuberculosis |
Inhibitors | Comment | Organism | Structure |
---|---|---|---|
3-nitropropionate | a succinate analogue. Inhibition by 3-nitropropionate proceeds through an unusual double slow-onset process featuring formation of a complex with a Ki of 0.0033 mM during the first minute, followed by formation of a final complex with a Ki* of 44 nM over the course of several min to hours | Mycobacterium tuberculosis | |
glyoxylate | - |
Mycobacterium tuberculosis | |
Itaconate | a succinate analogue lacking an acidic alpha-proton, the inhhibitor does not display slow-binding behavior | Mycobacterium tuberculosis |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
additional information | - |
additional information | stopped-flow spectroscopy and steady-state kinetics, solvent kinetic isotope effects, overview. Whereas the D2OV is consistent with partially rate-limiting proton transfer during formation of the hydroxyl group of isocitrate, the large inverse D2O(V/Ksuccinate) indicates that substantially different kinetic parameters exist when the enzyme is saturated with succinate | Mycobacterium tuberculosis | |
0.045 | - |
isocitrate | recombinant detagged enzyme, pH 7.0, 37°C | Mycobacterium tuberculosis | |
0.14 | - |
glyoxylate | recombinant detagged enzyme, pH 7.0, 37°C | Mycobacterium tuberculosis | |
0.412 | - |
succinate | recombinant detagged enzyme, pH 7.0, 37°C | Mycobacterium tuberculosis |
Metals/Ions | Comment | Organism | Structure |
---|---|---|---|
Mg2+ | required | Mycobacterium tuberculosis |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
isocitrate | Mycobacterium tuberculosis | - |
succinate + glyoxylate | - |
r | |
isocitrate | Mycobacterium tuberculosis H37R | - |
succinate + glyoxylate | - |
r |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Mycobacterium tuberculosis | - |
- |
- |
Mycobacterium tuberculosis H37R | - |
- |
- |
Purification (Comment) | Organism |
---|---|
recombinant N-terminally His6-tagged enzyme from Escherichia coli strain BL21(DE3) by nicke affinity chromatography, cleavage of the His-tag by thrombin, and dialysis | Mycobacterium tuberculosis |
Reaction | Comment | Organism | Reaction ID |
---|---|---|---|
isocitrate = succinate + glyoxylate | common mechanism for catalysis with succinate: (1) an unfavorable prebinding isomerization of the active site Cys191-His193 pair to the thiolate-imidazolium form, a process that is favored in D2O, and (2) the transfer of a proton from succinate or 3-NP to Cys191 | Mycobacterium tuberculosis |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
isocitrate | - |
Mycobacterium tuberculosis | succinate + glyoxylate | - |
r | |
isocitrate | - |
Mycobacterium tuberculosis H37R | succinate + glyoxylate | - |
r | |
succinate + glyoxylate | - |
Mycobacterium tuberculosis | isocitrate | - |
r | |
succinate + glyoxylate | - |
Mycobacterium tuberculosis H37R | isocitrate | - |
r |
Synonyms | Comment | Organism |
---|---|---|
ICL | - |
Mycobacterium tuberculosis |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
37 | - |
assay at | Mycobacterium tuberculosis |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
8.5 | - |
succinate | recombinant detagged enzyme, pH 7.0, 37°C | Mycobacterium tuberculosis | |
12.2 | - |
isocitrate | recombinant detagged enzyme, pH 7.0, 37°C | Mycobacterium tuberculosis |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
7 | - |
assay at | Mycobacterium tuberculosis |
Ki Value [mM] | Ki Value maximum [mM] | Inhibitor | Comment | Organism | Structure |
---|---|---|---|---|---|
additional information | - |
additional information | solvent isotope effects in inhibition by itaconate and 3-nitropropionate, overview | Mycobacterium tuberculosis | |
4.3 | - |
glyoxylate | recombinant detagged enzyme, pH 7.0, 37°C | Mycobacterium tuberculosis |
General Information | Comment | Organism |
---|---|---|
physiological function | the enzyme catalyzes the first committed step in the glyoxylate cycle, it is essential in Mycobacterium tuberculosis for cell survival during chronic infection | Mycobacterium tuberculosis |