Cloned (Comment) | Organism |
---|---|
gene glpX, sequence comparisons of class II FBPases, recombinant expression of His-tagged wild-type and mutant enzymes in Escherichia coli strain BL21(DE3) | Mycobacterium tuberculosis |
Protein Variants | Comment | Organism |
---|---|---|
T84A | site-directed mutagenesis, the codon ACC for Thr84 is replaced by GCA for alanine, the active site mutant shows fully abolished enzyme activity while retaining substrate binding affinity | Mycobacterium tuberculosis |
T84A | mutantion fully abolishes enzyme activity while retaining substrate binding affinity | Mycobacterium tuberculosis |
T84S | site-directed mutagenesis, the codon ACC for Thr84 is replaced by AGC for serine, the active site mutant retains some activity having a 10times reduction in Vmax and exhibit similar sensitivity to lithium when compared to the wild-type enzyme | Mycobacterium tuberculosis |
T84S | mutantion retains some activity having a 10 times reduction in Vmax and exhibits similar sensitivity to lithium when compared to the wild-type enzyme. Homology modeling using the Escherichia coli enzyme structure suggests that the replacement of the critical nucleophile OH- in the Thr84 residue of the wild-type enzyme by Ser84 results in subtle alterations of the position and orientation that reduces the catalytic efficiency. This mutant can be used to trap reaction intermediates, through crystallographic methods, facilitating the design of potent inhibitors via structure-based drug design | Mycobacterium tuberculosis |
Inhibitors | Comment | Organism | Structure |
---|---|---|---|
Li+ | - |
Mycobacterium tuberculosis | |
lithium | - |
Mycobacterium tuberculosis | |
additional information | enzyme mutant T84S can be used to trap reaction intermediates, through crystallographic methods, facilitating the design of potent inhibitors via structure-based drug design | Mycobacterium tuberculosis |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
additional information | - |
additional information | kinetics | Mycobacterium tuberculosis | |
0.0037 | - |
D-fructose 1,6-bisphosphate | pH 8.0, 22°C, recombinant wild-type enzyme | Mycobacterium tuberculosis | |
0.0057 | - |
D-fructose 1,6-bisphosphate | pH 8.0, 22°C, recombinant mutant T84S | Mycobacterium tuberculosis |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
D-fructose 1,6-bisphosphate + H2O | Mycobacterium tuberculosis | - |
D-fructose 6-phosphate + phosphate | - |
? | |
D-fructose 1,6-bisphosphate + H2O | Mycobacterium tuberculosis H37Rv | - |
D-fructose 6-phosphate + phosphate | - |
? | |
D-fructose 1,6-bisphosphate + H2O | Mycobacterium tuberculosis ATCC 25618 | - |
D-fructose 6-phosphate + phosphate | - |
? |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Mycobacterium tuberculosis | P9WN21 | - |
- |
Mycobacterium tuberculosis ATCC 25618 | P9WN21 | - |
- |
Mycobacterium tuberculosis H37Rv | P9WN21 | - |
- |
Purification (Comment) | Organism |
---|---|
- |
Mycobacterium tuberculosis |
recombinant His-tagged wild-type and mutant enzymes from Escherichia coli strain BL21(DE3) by nickel affinity chromatography, ultrafiltration, and desalting gel filtration | Mycobacterium tuberculosis |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
D-fructose 1,6-bisphosphate + H2O | - |
Mycobacterium tuberculosis | D-fructose 6-phosphate + phosphate | - |
? | |
D-fructose 1,6-bisphosphate + H2O | - |
Mycobacterium tuberculosis H37Rv | D-fructose 6-phosphate + phosphate | - |
? | |
D-fructose 1,6-bisphosphate + H2O | - |
Mycobacterium tuberculosis ATCC 25618 | D-fructose 6-phosphate + phosphate | - |
? | |
additional information | substrate binding analysis of wild-type and mutant enzymes by surface plasmon resonance method, substrate affinities, overview | Mycobacterium tuberculosis | ? | - |
? | |
additional information | substrate binding analysis of wild-type and mutant enzymes by surface plasmon resonance method, substrate affinities, overview | Mycobacterium tuberculosis H37Rv | ? | - |
? | |
additional information | substrate binding analysis of wild-type and mutant enzymes by surface plasmon resonance method, substrate affinities, overview | Mycobacterium tuberculosis ATCC 25618 | ? | - |
? |
Subunits | Comment | Organism |
---|---|---|
? | x * 36584, recombinant mutant T84A, mass spectrometry and sequence calculation, x * 36599, recombinant mutant T84S, mass spectrometry and sequence calculation | Mycobacterium tuberculosis |
Synonyms | Comment | Organism |
---|---|---|
class II fructose-1,6-bisphosphatase | - |
Mycobacterium tuberculosis |
fructose-1,6-bisphosphatase | - |
Mycobacterium tuberculosis |
fructose-1,6-bisphosphatase class II | - |
Mycobacterium tuberculosis |
GlpX | - |
Mycobacterium tuberculosis |
MtFBPaseII | - |
Mycobacterium tuberculosis |
MtFPBase | - |
Mycobacterium tuberculosis |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
22 | - |
assay at room temperature | Mycobacterium tuberculosis |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.22 | - |
D-fructose 1,6-bisphosphate | pH 8.0, 22°C, recombinant mutant T84S | Mycobacterium tuberculosis | |
2.1 | - |
D-fructose 1,6-bisphosphate | pH 8.0, 22°C, recombinant wild-type enzyme | Mycobacterium tuberculosis |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
8 | - |
assay at | Mycobacterium tuberculosis |
IC50 Value | IC50 Value Maximum | Comment | Organism | Inhibitor | Structure |
---|---|---|---|---|---|
0.25 | - |
pH 8.0, 22°C, recombinant wild-type enzyme | Mycobacterium tuberculosis | Li+ | |
0.28 | - |
pH 8.0, 22°C, recombinant mutant T84S | Mycobacterium tuberculosis | Li+ |
General Information | Comment | Organism |
---|---|---|
malfunction | homology modeling using the Escherichia coli enzyme structure suggests that the replacement of the critical nucleophile OH- in the Thr84 residue of the wild-type MtFBPase by Ser84 results in subtle alterations of the position and orientation that reduce the catalytic efficiency | Mycobacterium tuberculosis |
metabolism | essential enzyme for pathogenesis | Mycobacterium tuberculosis |
additional information | Thr84 in MtFBPaseII protein (Thr90 in Escherichia coli) is a conserved residue in the active site as shown in other FBPases | Mycobacterium tuberculosis |
physiological function | class II fructose-1,6-bisphosphatase enzyme in Mycobacterium tuberculosis is an essential enzyme for pathogenesis | Mycobacterium tuberculosis |