Cloned (Comment) | Organism |
---|---|
TcPRACA is expressed in Escherichia coli BL21 (DE3) | Trypanosoma cruzi |
Crystallization (Comment) | Organism |
---|---|
best crystals are obtained by mixing 2-3 microliter of the protein solution with an equal volume of crystallization buffer (0.1 M ammonium acetate/50 mM tri-sodium citrate dihydrate, pH 5.6/15% w/v polyethylene glycol 4000), equilibrated over 1 ml of the same buffer | Trypanosoma cruzi |
crystal structure analysis shows that TcPRACA is a homodimer, with each monomer folded in two symmetric alpha/beta subunits separated by a deep crevice. The structure of TcPRACA in complex with a transition-state analog, pyrrole-2-carboxylic acid, reveals the presence of one reaction center per monomer, with two Cys residues optimally located to perform acid/base catalysis through a carbanion stabilization mechanism | Trypanosoma cruzi |
Protein Variants | Comment | Organism |
---|---|---|
C130S | mutation of the catalytic Cys residues abolishes the enzymatic activity but preserves the mitogenic properties of the protein | Trypanosoma cruzi |
C300S | mutation of the catalytic Cys residues abolishes the enzymatic activity but preserves the mitogenic properties of the protein | Trypanosoma cruzi |
Inhibitors | Comment | Organism | Structure |
---|---|---|---|
pyrrole-2-carboxylic acid | PYC, competitive inhibitor | Trypanosoma cruzi |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Trypanosoma cruzi | Q4DA80 | - |
- |
Purification (Comment) | Organism |
---|---|
TcPRACA protein is purified with immobilized metal affinity chromatography on nickel columns. The active peak is further submitted to gel filtration chromatography at 2.5 ml/min in a Superdex200 26/60 column | Trypanosoma cruzi |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
L-proline | - |
Trypanosoma cruzi | D-proline | - |
? |
Subunits | Comment | Organism |
---|---|---|
homodimer | chrystal structure analysis | Trypanosoma cruzi |
Synonyms | Comment | Organism |
---|---|---|
proline racemase | - |
Trypanosoma cruzi |
TcPRACA | - |
Trypanosoma cruzi |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
37 | - |
assay at | Trypanosoma cruzi |