Crystal structure, catalytic mechanism, and mitogenic properties of Trypanosoma cruzi proline racemase
10.1073/pnas.0509010103
Crossref journal-article
Proceedings of the National Academy of Sciences
Proceedings of the National Academy of Sciences (341)
Abstract

Amino acid racemases catalyze the stereoinversion of the chiral C α to produce the d -enantiomers that participate in biological processes, such as cell wall construction in prokaryotes. Within this large protein family, bacterial proline racemases have been extensively studied as a model of enzymes acting with a pyridoxal-phosphate-independent mechanism. Here we report the crystal structure of the proline racemase from the human parasite Trypanosoma cruzi ( Tc PRACA), a secreted enzyme that triggers host B cell polyclonal activation, which prevents specific humoral immune responses and is crucial for parasite evasion and fate. The enzyme is a homodimer, with each monomer folded in two symmetric α/β subunits separated by a deep crevice. The structure of Tc PRACA 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. Mutation of the catalytic Cys residues abolishes the enzymatic activity but preserves the mitogenic properties of the protein. In contrast, inhibitor binding promotes the closure of the interdomain crevice and completely abrogates B cell proliferation, suggesting that the mitogenic properties of Tc PRACA depend on the exposure of transient epitopes in the ligand-free enzyme.

Bibliography

Buschiazzo, A., Goytia, M., Schaeffer, F., Degrave, W., Shepard, W., Grégoire, C., Chamond, N., Cosson, A., Berneman, A., Coatnoan, N., Alzari, P. M., & Minoprio, P. (2006). Crystal structure, catalytic mechanism, and mitogenic properties of Trypanosoma cruzi proline racemase. Proceedings of the National Academy of Sciences, 103(6), 1705–1710.

Authors 12
  1. Alejandro Buschiazzo (first)
  2. Maira Goytia (additional)
  3. Francis Schaeffer (additional)
  4. Wim Degrave (additional)
  5. William Shepard (additional)
  6. Christophe Grégoire (additional)
  7. Nathalie Chamond (additional)
  8. Alain Cosson (additional)
  9. Armand Berneman (additional)
  10. Nicolas Coatnoan (additional)
  11. Pedro M. Alzari (additional)
  12. Paola Minoprio (additional)
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Dates
Type When
Created 19 years, 7 months ago (Jan. 30, 2006, 9:03 p.m.)
Deposited 3 years, 2 months ago (June 7, 2022, 2:15 a.m.)
Indexed 2 weeks ago (Aug. 21, 2025, 12:53 p.m.)
Issued 19 years, 7 months ago (Jan. 30, 2006)
Published 19 years, 7 months ago (Jan. 30, 2006)
Published Online 19 years, 7 months ago (Jan. 30, 2006)
Published Print 19 years, 6 months ago (Feb. 7, 2006)
Funders 0

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@article{Buschiazzo_2006, title={Crystal structure, catalytic mechanism, and mitogenic properties of Trypanosoma cruzi proline racemase}, volume={103}, ISSN={1091-6490}, url={http://dx.doi.org/10.1073/pnas.0509010103}, DOI={10.1073/pnas.0509010103}, number={6}, journal={Proceedings of the National Academy of Sciences}, publisher={Proceedings of the National Academy of Sciences}, author={Buschiazzo, Alejandro and Goytia, Maira and Schaeffer, Francis and Degrave, Wim and Shepard, William and Grégoire, Christophe and Chamond, Nathalie and Cosson, Alain and Berneman, Armand and Coatnoan, Nicolas and Alzari, Pedro M. and Minoprio, Paola}, year={2006}, month=jan, pages={1705–1710} }