Crystallization (Comment) | Organism |
---|---|
mutant enzyme R191S, hanging drop vapor diffusion method, crystal structure of the R191S mutant complexed with 2-phosphoglycolate is determined at 1.65 A resolution | Trypanosoma brucei |
Protein Variants | Comment | Organism |
---|---|---|
D227A | mutant enzyme with reduced stability to 3.2 M guanidinium-HCl and reduced thermal stability at 42°C. The ratio of turnover number to Km for D-glyceraldehyde 3-phosphate as substrate is 2.2fold lower than the value for the wild-type enzyme. The ratio of turnover number to Km for glycerone phosphate as substrate is 5fold lower than the value for the wild-type enzyme | Trypanosoma brucei |
D227N | mutant enzyme with reduced stability to 3.2 M guanidinium-HCl and reduced thermal stability at 42°C. The ratio of turnover number to Km for D-glyceraldehyde 3-phosphate as substrate is 3.75fold lower than the value for the wild-type enzyme. The ratio of turnover number to Km for glycerone phosphate as substrate is 3.5fold lower than the value for the wild-type enzyme | Trypanosoma brucei |
R191A | mutant enzyme with reduced stability to 3.2 M guanidinium-HCl and reduced thermal stability at 42°C. The ratio of turnover number to Km for D-glyceraldehyde 3-phosphate as substrate is 10.4fold lower than the value for the wild-type enzyme. The ratio of turnover number to Km for glycerone phosphate as substrate is 10.6fold lower than the value for the wild-type enzyme | Trypanosoma brucei |
R191S | mutant enzyme with reduced stability to 3.2 M guanidinium-HCl and reduced thermal stability at 42°C. The ratio of turnover number to Km for D-glyceraldehyde 3-phosphate as substrate is 2fold lower than the value for the wild-type enzyme. The ratio of turnover number to Km for glycerone phosphate as substrate is 6.7fold lower than the value for the wild-type enzyme | Trypanosoma brucei |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.24 | - |
D-glyceraldehyde 3-phosphate | mutant enzyme S227A | Trypanosoma brucei | |
0.25 | - |
D-glyceraldehyde 3-phosphate | wild-type enzyme | Trypanosoma brucei | |
0.28 | - |
D-glyceraldehyde 3-phosphate | mutant enzyme S227N | Trypanosoma brucei | |
0.34 | - |
D-glyceraldehyde 3-phosphate | mutant enzyme R191S | Trypanosoma brucei | |
0.67 | - |
glycerone phosphate | mutant enzyme R191A | Trypanosoma brucei | |
0.7 | - |
glycerone phosphate | mutant enzyme D227N | Trypanosoma brucei | |
0.9 | - |
glycerone phosphate | mutant enzyme D227A | Trypanosoma brucei | |
1 | - |
glycerone phosphate | wild-type enzyme | Trypanosoma brucei | |
1.1 | - |
D-glyceraldehyde 3-phosphate | mutant enzyme R191A | Trypanosoma brucei | |
1.6 | - |
glycerone phosphate | mutant enzyme R191S | Trypanosoma brucei |
Organic Solvent | Comment | Organism |
---|---|---|
guanidine-HCl | 3.2 M, half-life of wild-type enzyme: 30 min, half-life of mutant enzyme D227N: 20 min, half-life of mutant enzyme D227A: 15 min, half-life of mutant enzyme R191S: 8 min, half-life of mutant enzyme R191A: 19 min | Trypanosoma brucei |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Trypanosoma brucei | P04789 | brucei TIM | - |
Renatured (Comment) | Organism |
---|---|
protein unfolded in 3.2 M guanidinium-HCl is diluted to final concentrations of 0.32 M guanidinium-HCl, 16% recovery of wild-type enzyme after 72 h, 2.1-2.6% recovery of the mutant enzymes D227N, D227A, R191S and R191A after 72 h | Trypanosoma brucei |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
D-Glyceraldehyde 3-phosphate | - |
Trypanosoma brucei | Glycerone phosphate | - |
r |
Temperature Stability Minimum [°C] | Temperature Stability Maximum [°C] | Comment | Organism |
---|---|---|---|
42 | - |
half-life of wild-type enzyme: 180 min, half-life of mutant enzyme D227N: 128 min, half-life of mutant enzyme D227A: 122 min, half-life of mutant enzyme R191S: 87 min, half-life of mutant enzyme R191A: 124 min | Trypanosoma brucei |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
63 | - |
glycerone phosphate | mutant enzyme R191A | Trypanosoma brucei | |
180 | - |
glycerone phosphate | mutant enzyme D227A | Trypanosoma brucei | |
200 | - |
glycerone phosphate | mutant enzyme D227N | Trypanosoma brucei | |
240 | - |
glycerone phosphate | mutant enzyme R191S | Trypanosoma brucei | |
1000 | - |
glycerone phosphate | wild-type enzyme | Trypanosoma brucei | |
1600 | - |
D-glyceraldehyde 3-phosphate | mutant enzyme S227N | Trypanosoma brucei | |
2500 | - |
D-glyceraldehyde 3-phosphate | mutant enzymeR191A | Trypanosoma brucei | |
2600 | - |
D-glyceraldehyde 3-phosphate | mutant enzyme S227A | Trypanosoma brucei | |
4000 | - |
D-glyceraldehyde 3-phosphate | mutant enzyme R191S | Trypanosoma brucei | |
6000 | - |
D-glyceraldehyde 3-phosphate | wild-type enzyme | Trypanosoma brucei |