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Literature summary for 4.1.3.1 extracted from
- Mycobacterium tuberculosis isocitrate lyase (MtbIcl): role of divalent cations in modulation of functional and structural properties (2008), Proteins, 72, 892-900.
Cloned(Commentary)
| Cloned (Comment) | Organism |
|---|---|
| from genomic DNA from strain H37Rv in pET21d for inducible expression in Escherichia coli C41 | Mycobacterium tuberculosis |
Inhibitors
| Inhibitors | Comment | Organism | Structure |
|---|---|---|---|
| CdCl2 | - |
Mycobacterium tuberculosis |  |
| ZnCl2 | - |
Mycobacterium tuberculosis | |
Metals/Ions
| Metals/Ions | Comment | Organism | Structure |
|---|---|---|---|
| Cd2+ | 5 mM, 90% inhibition of Mg2+-activated enzyme, interaction with catalytic domain induces partial unfolding (revealed by thermal denaturation, far-UV circular dichroism, fluorescence spectra, and glutaraldehyde crosslinking) | Mycobacterium tuberculosis | |
| Mg2+ | 5 mM, essential for catalytic activity, 100% activity, binds and stabilizes non-catalytic alpha/beta barrel core without structural alterations (revealed by thermal, urea and GdnHCL denaturation and far-UV CD) | Mycobacterium tuberculosis | |
| Mn2+ | 5 mM, 45% activity, binds and stabilizes non-catalytic alpha/beta barrel core without structural alterations (revealed by thermal, urea and GdnHCL denaturation and far-UV CD) | Mycobacterium tuberculosis | |
| Zn2+ | 5 mM, 90% inhibition of Mg2+-activated enzyme, interaction with catalytic domain induces partial unfolding (revealed by thermal denaturation, far-UV circular dichroism, ANS fluorescence spectra, and glutaraldehyde crosslinking) | Mycobacterium tuberculosis | |
Molecular Weight [Da]
| Molecular Weight [Da] | Molecular Weight Maximum [Da] | Comment | Organism |
|---|---|---|---|
| 48000 | - |
single peak in ESI-MS | Mycobacterium tuberculosis |
| 200000 | - |
gel filtration | Mycobacterium tuberculosis |
Natural Substrates/ Products (Substrates)
| Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
|---|---|---|---|---|---|---|
| (1S,2S)-1-hydroxypropane-1,2,3-tricarboxylate | Mycobacterium tuberculosis | reversible aldol cleavage | succinate + glyoxylate | glyoxylate shunt | r | |
Organism
| Organism | UniProt | Comment | Textmining |
|---|---|---|---|
| Mycobacterium tuberculosis | - |
- |
- |
Purification (Commentary)
| Purification (Comment) | Organism |
|---|---|
| recombinant protein from cell lysate by Ni-NTA metal-affinity chromatography (elution: 400 mM imidazole) and gel filtration on Superdex 200HR 10/300 column | Mycobacterium tuberculosis |
Substrates and Products (Substrate)
| Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
|---|---|---|---|---|---|---|
| (1S,2S)-1-hydroxypropane-1,2,3-tricarboxylate | reversible aldol cleavage | Mycobacterium tuberculosis | succinate + glyoxylate | glyoxylate shunt | r | |
| isocitrate | 30°C, pH 7.5, HEPES containing phenylhydrazine HCl | Mycobacterium tuberculosis | succinate + glyoxylate | absorbance of glyoxylate phenylhydrazone at 334 nm | r | |
Subunits
| Subunits | Comment | Organism |
|---|---|---|
| tetramer | 4 * 48000, gel filtration | Mycobacterium tuberculosis |
Synonyms
| Synonyms | Comment | Organism |
|---|---|---|
| isocitrate lyase | - |
Mycobacterium tuberculosis |
| MtbIcl | - |
Mycobacterium tuberculosis |
Temperature Stability [°C]
| Temperature Stability Minimum [°C] | Temperature Stability Maximum [°C] | Comment | Organism |
|---|---|---|---|
| 50 | - |
5 mM ZnCl2, sigmoidal thermal denaturation curve, loss of 35% of helicity | Mycobacterium tuberculosis |
| 51 | - |
native enzyme, apparent melting temperature (Tm), far-UV CD (222 nm), major part of enzyme resistant to thermal unfolding despite almost complete loss of activity above 50°C, catalytic domain unfolded | Mycobacterium tuberculosis |
| 55 | - |
2.5 mM MgCl2, first Tm of biphasic thermal denaturation curve (far-UV CD (222 nm)), loss of secondary structure similar to native enzyme, catalytic domain unfolded | Mycobacterium tuberculosis |
| 56 | - |
1 mM MgCl2, first Tm of biphasic thermal denaturation curve (far-UV CD (222 nm)), loss of secondary structure similar to native enzyme, catalytic domain unfolded | Mycobacterium tuberculosis |
| 66 | - |
2.5 mM MgCl2, second Tm of biphasic thermal denaturation curve (far-UV CD (222 nm)), almost complete loss of secondary structure, noncatalytic alpha/beta barrel unfolded | Mycobacterium tuberculosis |
| 90 | - |
1 mM MgCl2, second Tm of biphasic thermal denaturation curve (far-UV CD (222 nm)), almost complete loss of secondary structure, noncatalytic alpha/beta barrel unfolded | Mycobacterium tuberculosis |
IC50 Value
| IC50 Value | IC50 Value Maximum | Comment | Organism | Inhibitor | Structure |
|---|---|---|---|---|---|
| 0.6 | - |
- |
Mycobacterium tuberculosis | CdCl2 | |
| 0.8 | - |
- |
Mycobacterium tuberculosis | ZnCl2 | |
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