Cloned (Comment) | Organism |
---|---|
expression of wild-type and mutant enzymes in TIM-deficient Escherichia coli strain AA200 | Plasmodium falciparum |
Crystallization (Comment) | Organism |
---|---|
wild-type and mutant enzymes in complex with inhibitor 3-phosphoglycerate, 0.003 ml of 10 mg/ml protein in 100 mM triethanolamine-HCl, pH 7.6, are mixed with an equal volume of reservoir solution, the crystallization cocktail contains 0.1 M sodium acetate pH 4.0-5.5 and PEG 1450 varying from 8% to 24% in the reservoir, X-ray diffraction structure determination and analysis at 1.4-2.25 A resolutions | Plasmodium falciparum |
Protein Variants | Comment | Organism |
---|---|---|
F96H | site-directed mutagenesis, the mutant exhibits highly reduced catalytic efficiency and decreased substrate-binding affinity, as well as reduced sensitivity to inhibitor 3-phosphoglycerate, compared to the wild-type enzyme | Plasmodium falciparum |
F96S | site-directed mutagenesis, the mutant exhibits highly reduced catalytic efficiency and decreased substrate-binding affinity, as well as reduced sensitivity to inhibitor 3-phosphoglycerate, compared to the wild-type enzyme | Plasmodium falciparum |
F96W | site-directed mutagenesis, the mutant exhibits reduced catalytic efficiency and decreased substrate-binding affinity, as well as reduced sensitivity to inhibitor 3-phosphoglycerate, compared to the wild-type enzyme. The wild-type enzyme shows a loop-open state for 3-phosphoglycerate binding at the active site, while the mutant F96W shows both open and closed states | Plasmodium falciparum |
additional information | decrease in ligand affinity in F96 mutants can be a consequence of differences in the water network connecting residue 96 to Ser73 in the vicinity of the active site | Plasmodium falciparum |
Inhibitors | Comment | Organism | Structure |
---|---|---|---|
3-phosphoglycerate | binding at the active site with the dimer-interface site showing strong electrostatic anchoring of the phosphate group involving the Arg98 and Lys112 residues of TIM, comparisons of binding structures at the interface, overview | Plasmodium falciparum |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.39 | - |
D-glyceraldehyde 3-phosphate | pH 7.6, wild-type enzyme | Plasmodium falciparum | |
1.2 | - |
D-glyceraldehyde 3-phosphate | pH 7.6, mutant F96W | Plasmodium falciparum | |
2.18 | - |
D-glyceraldehyde 3-phosphate | pH 7.6, mutant F96S | Plasmodium falciparum | |
2.62 | - |
D-glyceraldehyde 3-phosphate | pH 7.6, mutant F96H | Plasmodium falciparum |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
D-Glyceraldehyde 3-phosphate | Plasmodium falciparum | - |
Glycerone phosphate | - |
? |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Plasmodium falciparum | Q07412 | - |
- |
Purification (Comment) | Organism |
---|---|
recombinant wild-type and mutant enzymes from TIM-deficient Escherichia coli strain AA200 by gel filtration and ion-exchange chromatography | Plasmodium falciparum |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
D-Glyceraldehyde 3-phosphate | - |
Plasmodium falciparum | Glycerone phosphate | - |
? | |
additional information | Plasmodium falciparum TIM is unique in possessing a Phe residue at position 96 in place of the conserved Ser that is found in TIMs from the majority of other organisms | Plasmodium falciparum | ? | - |
? |
Synonyms | Comment | Organism |
---|---|---|
TIM | - |
Plasmodium falciparum |
Triosephosphate isomerase | - |
Plasmodium falciparum |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
185 | - |
D-glyceraldehyde 3-phosphate | pH 7.6, mutant F96H | Plasmodium falciparum | |
620 | - |
D-glyceraldehyde 3-phosphate | pH 7.6, mutant F96S | Plasmodium falciparum | |
2466 | - |
D-glyceraldehyde 3-phosphate | pH 7.6, mutant F96W | Plasmodium falciparum | |
42330 | - |
D-glyceraldehyde 3-phosphate | pH 7.6, wild-type enzyme | Plasmodium falciparum |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
7.6 | - |
assay at | Plasmodium falciparum |
Ki Value [mM] | Ki Value maximum [mM] | Inhibitor | Comment | Organism | Structure |
---|---|---|---|---|---|
1.9 | - |
3-phosphoglycerate | pH 7.6, wild-type enzyme | Plasmodium falciparum | |
5 | - |
3-phosphoglycerate | above, pH 7.6, mutants F96W, F96S, and F96H | Plasmodium falciparum |
kcat/KM Value [1/mMs-1] | kcat/KM Value Maximum [1/mMs-1] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
28.5 | - |
D-glyceraldehyde 3-phosphate | pH 7.6, mutant F96S | Plasmodium falciparum | |
70.5 | - |
D-glyceraldehyde 3-phosphate | pH 7.6, mutant F96H | Plasmodium falciparum | |
2050 | - |
D-glyceraldehyde 3-phosphate | pH 7.6, mutant F96W | Plasmodium falciparum | |
10850 | - |
D-glyceraldehyde 3-phosphate | pH 7.6, wild-type enzyme | Plasmodium falciparum |