Cloned (Comment) | Organism |
---|---|
into pQE30 vector. N-terminally MRGS(H)6GS-tagged GloII wild-type and mutant enzymes transformed into Escherichia coli competent XL1-Blue cells or EZ cells | Plasmodium falciparum |
Protein Variants | Comment | Organism |
---|---|---|
K260D | unaltered metal binding sites and is correctly folded, but reduced activity. Has a much lower catalytic efficiency, but the spectrum is very similar to the wild-type enzyme. Has a highly simplified bell-shaped pH-activity profile with a maximum at pH 6.7 in contrast to the broad asymmetric profiles of the other enzymes | Plasmodium falciparum |
K260Q | unaltered metal binding sites and is correctly folded, but reduced activity | Plasmodium falciparum |
R154K | unaltered metal binding sites and is correctly folded, but reduced activity | Plasmodium falciparum |
R154M | unaltered metal binding sites and is correctly folded, but reduced activity | Plasmodium falciparum |
R257D | unaltered metal binding sites and is correctly folded, but reduced activity | Plasmodium falciparum |
R257Q | unaltered metal binding sites and is correctly folded, but reduced activity | Plasmodium falciparum |
Inhibitors | Comment | Organism | Structure |
---|---|---|---|
D-lactate | competitive inhibition | Plasmodium falciparum | |
glutathione | non-competitive inhibition. Is a stronger inhibitor than the ethyl ester of glutathione. Theorell-Chance Bi Bi mechanism with glutathione as the second product | Plasmodium falciparum | |
glutathione ethyl ester | carboxylate group of the glycine-moiety is modified, non-competitive inhibition | Plasmodium falciparum |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.114 | - |
S-D-lactoylglutathione | wild-type, at 25°C, in 100 mM MOPS/NaOH, pH 6.8 | Plasmodium falciparum | |
0.116 | - |
S-D-lactoylglutathione | mutant R154K, at 25°C, in 100 mM MOPS/NaOH, pH 6.8 | Plasmodium falciparum | |
0.275 | - |
S-D-lactoylglutathione | mutant R154M, at 25°C, in 100 mM MOPS/NaOH, pH 6.8 | Plasmodium falciparum | |
2.551 | - |
S-D-lactoylglutathione | mutant R257Q, at 25°C, in 100 mM MOPS/NaOH, pH 7.4 | Plasmodium falciparum | |
2.867 | - |
S-D-lactoylglutathione | mutant K260D, at 25°C, in 100 mM MOPS/NaOH, pH 6.8 | Plasmodium falciparum | |
4.308 | - |
S-D-lactoylglutathione | mutant K260Q, at 25°C, in 100 mM MOPS/NaOH, pH 6.8 | Plasmodium falciparum | |
5.782 | - |
S-D-lactoylglutathione | mutant R257D, at 25°C, in 100 mM MOPS/NaOH, pH 6.8 | Plasmodium falciparum |
Metals/Ions | Comment | Organism | Structure |
---|---|---|---|
Zinc | recombinant GloII is zinc-dependent | Plasmodium falciparum |
Molecular Weight [Da] | Molecular Weight Maximum [Da] | Comment | Organism |
---|---|---|---|
31800 | - |
x * 31800, His-tagged GloII, SDS-PAGE | Plasmodium falciparum |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Plasmodium falciparum | - |
- |
- |
Purification (Comment) | Organism |
---|---|
wild-type and glutathione-binding site mutants, by sonication, centrifugation and on Ni-NTA column | Plasmodium falciparum |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
S-D-lactoylglutathione + H2O | substrate is predominantly bound via ionic interactions with the conserved residues Arg257 and Lys260. Correct substrate binding is a pH- and salt-dependent rate-limiting step for catalysis. Bi Bi mechanism with the direct involvement of a hydroxide ion as a second substrate | Plasmodium falciparum | D-lactate + glutathione | - |
? |
Subunits | Comment | Organism |
---|---|---|
? | x * 31800, His-tagged GloII, SDS-PAGE | Plasmodium falciparum |
Synonyms | Comment | Organism |
---|---|---|
GloII | - |
Plasmodium falciparum |
glyoxalase II | - |
Plasmodium falciparum |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
60 | - |
S-D-lactoylglutathione | mutant R257D, at 25°C, in 100 mM MOPS/NaOH, pH 6.8 | Plasmodium falciparum | |
67 | - |
S-D-lactoylglutathione | mutant R257Q, at 25°C, in 100 mM MOPS/NaOH, pH 7.4 | Plasmodium falciparum | |
74 | - |
S-D-lactoylglutathione | mutant K260D, at 25°C, in 100 mM MOPS/NaOH, pH 6.8 | Plasmodium falciparum | |
171 | - |
S-D-lactoylglutathione | mutant K260Q, at 25°C, in 100 mM MOPS/NaOH, pH 6.8 | Plasmodium falciparum | |
222 | - |
S-D-lactoylglutathione | mutant R154M, at 25°C, in 100 mM MOPS/NaOH, pH 6.8 | Plasmodium falciparum | |
285 | - |
S-D-lactoylglutathione | mutant R154K, at 25°C, in 100 mM MOPS/NaOH, pH 6.8 | Plasmodium falciparum | |
375 | - |
S-D-lactoylglutathione | wild-type, at 25°C, in 100 mM MOPS/NaOH, pH 6.8 | Plasmodium falciparum |
Ki Value [mM] | Ki Value maximum [mM] | Inhibitor | Comment | Organism | Structure |
---|---|---|---|---|---|
0.000031 | - |
D-lactate | inhibitor binding to the free enzyme, at 25°C, pH 6.8 | Plasmodium falciparum | |
3.5 | - |
glutathione | inhibitor binding to the free enzyme, at 25°C, pH 6.8 | Plasmodium falciparum | |
13 | - |
glutathione | inhibitor binding to the occupied enzyme, at 25°C, pH 6.8 | Plasmodium falciparum | |
14 | - |
glutathione ethyl ester | inhibitor binding to the free enzyme, at 25°C, pH 6.8 | Plasmodium falciparum | |
49 | - |
glutathione ethyl ester | inhibitor binding to the occupied enzyme, at 25°C, pH 6.8 | Plasmodium falciparum |
kcat/KM Value [1/mMs-1] | kcat/KM Value Maximum [1/mMs-1] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
11 | - |
S-D-lactoylglutathione | mutant R257D, at 25°C, in 100 mM MOPS/NaOH, pH 6.8 | Plasmodium falciparum | |
26 | - |
S-D-lactoylglutathione | mutant K260D, at 25°C, in 100 mM MOPS/NaOH, pH 6.8 | Plasmodium falciparum | |
30 | - |
S-D-lactoylglutathione | mutant R257Q, at 25°C, in 100 mM MOPS/NaOH, pH 7.4 | Plasmodium falciparum | |
41 | - |
S-D-lactoylglutathione | mutant K260Q, at 25°C, in 100 mM MOPS/NaOH, pH 6.8 | Plasmodium falciparum | |
810 | - |
S-D-lactoylglutathione | mutant R154M, at 25°C, in 100 mM MOPS/NaOH, pH 6.8 | Plasmodium falciparum | |
2400 | - |
S-D-lactoylglutathione | mutant R154K, at 25°C, in 100 mM MOPS/NaOH, pH 6.8 | Plasmodium falciparum | |
3300 | - |
S-D-lactoylglutathione | wild-type, at 25°C, in 100 mM MOPS/NaOH, pH 6.8 | Plasmodium falciparum |