Literature summary for 5.1.1.4 extracted from

Buschiazzo, A.; Goytia, M.; Schaeffer, F.; Degrave, W.; Shepard, W.; Gregoire, C.; Chamond, N.; Cosson, A.; Berneman, A.; Coatnoan, N.; Alzari, P.M.; Minoprio, P.
Crystal structure, catalytic mechanism, and mitogenic properties of Trypanosoma cruzi proline racemase (2006), Proc. Natl. Acad. Sci. USA, 103, 1705-1710.

Cloned(Commentary)

Cloned (Comment)	Organism
TcPRACA is expressed in Escherichia coli BL21 (DE3)	Trypanosoma cruzi

Crystallization (Commentary)

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

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

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

Inhibitors	Comment	Organism	Structure
pyrrole-2-carboxylic acid	PYC, competitive inhibitor	Trypanosoma cruzi

Organism

Organism	UniProt	Comment	Textmining
Trypanosoma cruzi	Q4DA80	-	-

Purification (Commentary)

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 and Products (Substrate)

Substrates	Comment Substrates	Organism	Products	Comment (Products)	Rev.	Reac.
L-proline	-	Trypanosoma cruzi	D-proline	-	?

Subunits

Subunits	Comment	Organism
homodimer	chrystal structure analysis	Trypanosoma cruzi

Synonyms

Synonyms	Comment	Organism
proline racemase	-	Trypanosoma cruzi
TcPRACA	-	Trypanosoma cruzi

Temperature Optimum [°C]

Temperature Optimum [°C]	Temperature Optimum Maximum [°C]	Comment	Organism
37	-	assay at	Trypanosoma cruzi